Oxysterols and methods of use thereof

ABSTRACT

Compounds are provided according to Formula (A): and pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof; wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and R G  are as defined herein. Compounds of the present invention are contemplated useful for the prevention and treatment of a variety of conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a U.S. National Phase Application under 35U.S.C. § 371 of International Application Number PCT/US2017/057276,filed Oct. 18, 2017, which claims priority to and the benefit of U.S.Provisional Application No. 62/409,756, filed Oct. 18, 2016, and U.S.Provisional Application No. 62/409,768, filed Oct. 18, 2016, each ofwhich are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

NMDA receptors are heteromeric complexes comprised of NR1, NR2, and/orNR3 subunits and possess distinct recognition sites for exogenous andendogenous ligands. These recognition sites include binding sites forglycine, and glutamate agonists and modulators. NMDA receptors areexpressed in the peripheral tissues and the CNS, where they are involvedin excitatory synaptic transmission. Activating these receptorscontributes to synaptic plasticity in some circumstances andexcitotoxicity in others. These receptors are ligand-gated ion channelsthat admit Ca²⁺ after binding of the glutamate and glycine, and arefundamental to excitatory neurotransmission and normal CNS function.Positive modulators may be useful as therapeutic agents with potentialclinical uses as cognitive enhancers and in the treatment of psychiatricdisorders in which glutamatergic transmission is reduced or defective(see, e.g., Horak et al., J. of Neuroscience, 2004, 24(46),10318-10325). In contrast, negative modulators may be useful astherapeutic agents with potential clinical uses in the treatment ofpsychiatric disorders in which glutamatergic transmission ispathologically increased (e.g., treatment resistant depression).

Oxysterols are cholesterol analogs that are modulators of NMDA receptorfunction. There is a need for new oxysterols that modulate the NMDAreceptor for the prevention and treatment of conditions associated withNMDA expression and function. Compounds, compositions, and methodsdescribed herein are directed toward this end.

SUMMARY OF THE INVENTION

Provided herein are substituted oxysterols useful for preventing and/ortreating a broad range of disorders, including, but not limited to,NMDA-mediated disorders. Further provided are pharmaceuticalcompositions comprising the compounds of the present invention, andmethods of their use and treatment.

In one aspect, provided herein are compounds according to Formula (A):

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is hydrogen or alkyl (e.g., C₁-C₆ alkyl); each of R² and R³ isindependently hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R² and R³, togetherwith the carbon atom to which they are attached form a 3-8 memberedring; each of R⁴ and R⁵ is independently hydrogen, halo, or —OR^(C),wherein R^(C) is hydrogen or C₁-C₆ alkyl (e.g., C₁-C₃ alkyl), or R⁴ andR⁵, together with the carbon atom to which they are attached form an oxogroup; R⁶ is absent or hydrogen; R^(G) is hydrogen or alkyl; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl (e.g., C₁-C₆ alkyl). In someembodiments, R¹ is C₁-C₆ alkyl (e.g., —CH₃, —CH₂CH₃, —CH₂OCH₃, or —CF₃).In some embodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments,R¹ is —CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., C₁-C₃ alkyl).

In some embodiments, R² is hydrogen or alkyl (e.g., C₁-C₆ alkyl).

In some embodiments, each of R² and R³ is independently hydrogen oralkyl (e.g., C₁-C₆ alkyl). In some embodiments, each of R² and R³ isindependently hydrogen or C₁-C₆ haloalkyl (e.g., —CF₃). In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.

In some embodiments, R⁴ is —OH or halo (e.g., —F).

In some embodiments, R⁴ and R⁵, together with the carbon atom to whichthey are attached form an oxo group. In some embodiments, R⁴ is hydrogenand R⁵ is halo (e.g., —F). In some embodiments, R⁴ and R⁵ are halo(e.g., —F). In some embodiments, R⁴ and R⁵ are hydrogen.

In some embodiments, R² is aryl or heteroaryl and R³ is hydrogen. Insome embodiments, R² is carbocyclyl or heterocyclyl and R³ is hydrogen.In some embodiments, R² and R³ are hydrogen. In some embodiments, R² andR³, together with the carbon atom to which they are attached form a 3-8membered carbocyclic or heterocyclic ring.

In some embodiments, R⁶ is hydrogen and

represents a single bond.

In some embodiments, R^(G) is hydrogen or —CH₃.

In one aspect, provided herein are compounds according to Formula(I-63):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is alkyl(e.g., C₁-C₆ alkyl); each of R² and R³ is independently hydrogen, alkyl(e.g., C₁-C₆ alkyl), alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,or heteroaryl, or R² and R³, together with the carbon atom to which theyare attached form a 3-8 membered ring; each of R⁴ and R⁵ isindependently hydrogen, halo, or —OR^(C), wherein R^(C) is hydrogen orC₁-C₆ alkyl (e.g., C₁-C₃ alkyl), or R⁴ and R⁵, together with the carbonatom to which they are attached form an oxo group; R⁶ is absent orhydrogen; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl (e.g., C₁-C₆ alkyl). In someembodiments, R¹ is C₁-C₆ alkyl (e.g., —CH₃, —CH₂CH₃, —CH₂OCH₃, or —CF₃).In some embodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments,R¹ is —CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., C₁-C₃ alkyl). Insome embodiments, R¹ is unsubstituted alkyl. In some embodiments R¹ is—CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., —CH₃).

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroarylor R² and R³, together with the carbon atom to which they are attachedform a 3-8 membered ring.

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), carbocyclyl, heterocyclyl, aryl, or heteroaryl or R² and R³,together with the carbon atom to which they are attached form a 3-8membered ring.

In some embodiments, R² is hydrogen or alkyl (e.g., C₁-C₆ alkyl).

In some embodiments, each of R² and R³ is independently hydrogen oralkyl (e.g., C₁-C₆ alkyl). In some embodiments, each of R² and R³ isindependently hydrogen or C₁-C₆ haloalkyl (e.g., —CF₃). In someembodiments, each of R² and R³ is independently C₅ alkyl (e.g.,substituted or unsubstituted isopentyl) or hydrogen. In someembodiments, each of R² and R³ is independently isopentyl (e.g.,substituted or unsubstituted isopentyl) or hydrogen. In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.

In some embodiments, R⁴ is —OH or halo (e.g., —F).

In some embodiments, R⁴ and R⁵, together with the carbon atom to whichthey are attached form an oxo group. In some embodiments, R⁴ is hydrogenand R⁵ is halo (e.g., —F). In some embodiments, R⁴ and R⁵ are halo(e.g., —F). In some embodiments, R⁴ and R⁵ are hydrogen.

In some embodiments, R² is aryl or heteroaryl and R³ is hydrogen. Insome embodiments, R² is carbocyclyl or heterocyclyl and R³ is hydrogen.In some embodiments, R² and R³ are hydrogen. In some embodiments, R² isisopentyl (e.g., substituted or unsubstituted isopentyl) and R³ ishydrogen. In some embodiments, R² is —CF₃ or —CH₃ and R³ is hydrogen or—CH₃. In some embodiments, R² and R³, together with the carbon atom towhich they are attached form a 3-8 membered carbocyclic or heterocyclicring.

In some embodiments, R¹ is —CH₃ or —CH₂CH₃, R² is isopentyl (e.g.,substituted or unsubstituted isopentyl), and R³ is hydrogen. In someembodiments, R¹ is —CH₃ or —CH₂CH₃, R² is unsubstituted isopentyl, andR³ is hydrogen.

In some embodiments, R² is unsubstituted C₁-C₆ alkyl or C₁-C₆ haloalkyl.In some embodiments, R² is unsubstituted C₁-C₆ alkyl. In someembodiments, R² is pyridyl. In some embodiments, each of R² is isopentyland R³ is hydrogen. In some embodiments, R² is —CF₃ and R³ is hydrogen.In some embodiments, R² is unsubstituted alkyl (e.g., unsubstitutedC₁-C₆ alkyl). In some embodiments, R² is carbocyclylalkyl. In someembodiments, R² is carbocyclylalkyl and R³ is hydrogen. In someembodiments, R² is aralkyl (e.g., benzyl). In some embodiments, R² isheterocyclylalkyl. In some embodiments, wherein R² is unsubstitutedC₁-C₆ alkyl, C₁-C₆ haloalkyl, carbocyclyl, carbocyclylalkyl, aralkyl, orheterocyclylalkyl.

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-A63), (I-B63), or (I-C63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-A63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-C63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-B63):

In some embodiments, the compound of Formula (I63) is selected from acompound of Formula (I-D63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-E63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-D-i63) or (I-D-ii63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-E-i63) or (I-E-ii63):

In

In one aspect, provided herein are compounds according to Formula(I-67):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogenor alkyl (e.g., C₁-C₆ alkyl); each of R² and R³ is independentlyhydrogen, alkyl (e.g., C₁-C₆ alkyl), carbocyclyl, heterocyclyl, aryl, orheteroaryl, or R² and R³, together with the carbon atom to which theyare attached form a 3-8 membered ring; each of R⁴ and R⁵ isindependently hydrogen, halo, or —OR^(C), wherein R^(C) is hydrogen orC₁-C₆ alkyl (e.g., C₁-C₃ alkyl), or R⁴ and R⁵, together with the carbonatom to which they are attached form an oxo group; R⁶ is absent orhydrogen; and

represents a single or double bond, wherein when one of

is a bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl. In some embodiments, R¹ isunsubstituted alkyl. In some embodiments, R¹ is C₁-C₆ alkyl. In someembodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments, R¹ is—CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., —CH₃).

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), carbocyclyl, heterocyclyl, aryl, or heteroaryl or R² and R³,together with the carbon atom to which they are attached form a 3-8membered ring.

In some embodiments, R² is hydrogen or alkyl. In some embodiments, eachof R² and R³ is independently hydrogen or alkyl. In some embodiments,each of R² and R³ is independently hydrogen or C₁-C₆ haloalkyl. In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.In some embodiments, R⁴ is —OH or halo. In some embodiments, R⁴ and R⁵,together with the carbon atom to which they are attached form an oxogroup. In some embodiments, R⁴ is hydrogen and R⁵ is halo. In someembodiments, R⁴ and R⁵ are halo. In some embodiments, R⁴ and R⁵ arehydrogen. In some embodiments, R² is aryl or heteroaryl and R³ ishydrogen. In some embodiments, R² is carbocyclyl or heterocyclyl and R³is hydrogen. In some embodiments, R² and R³ are hydrogen. In someembodiments, R² and R³, together with the carbon atom to which they areattached form a 3-8 membered carbocyclic or heterocyclic ring.

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-A67), (I-B67), or (I-C67):

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-A67):

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-C67):

In an aspect, provided herein are compounds of selected from the groupconsisting of:

In an aspect, provided herein is a pharmaceutical composition comprisinga compound described herein, or pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

In an aspect, provided herein is a method of inducing sedation oranesthesia comprising administering to a subject an effective amount ofa compound described herein, or pharmaceutically acceptable saltthereof, or pharmaceutical composition thereof.

In an aspect, provided herein is a method for treating or preventing adisorder described herein, comprising administering to a subject in needthereof an effective amount of a compound described herein, orpharmaceutically acceptable salt thereof, or pharmaceutical compositionthereof.

In some embodiments, the disorder is a metabolic disorder.

In some embodiments, the disorder is an autoimmune disorder.

In some embodiments, the disorder is rheumatoid arthritis, juvenileidiopathic arthritis, ankylosing spondylitis, psoriatic arthritis,Crohn's disease, ulcerative colitis, and plaque psoriasis.

In some embodiments, the disorder is a gastrointestinal (GI) disordere.g., constipation, irritable bowel syndrome (IBS), inflammatory boweldisease (IBD) (e.g., ulcerative colitis, Crohn's disease), structuraldisorders affecting the GI, anal disorders (e.g., hemorrhoids, internalhemorrhoids, external hemorrhoids, anal fissures, perianal abscesses,anal fistula), colon polyps, cancer, or colitis.

In some embodiments, the disorder is inflammatory bowel disease.

In some embodiments, the disorder is cancer, diabetes, or a sterolsynthesis disorder.

In an aspect, provided herein is a method for treating or preventing aCNS-related condition comprising administering to a subject in needthereof an effective amount of a compound described herein, orpharmaceutically acceptable salt thereof, or pharmaceutical compositionthereof. In some embodiments, the CNS-related condition is an adjustmentdisorder, anxiety disorder (including obsessive-compulsive disorder,posttraumatic stress disorder, and social phobia), cognitive disorder(including Alzheimer's disease and other forms of dementia (e.g.,frontotemporal dementia), dissociative disorder, eating disorder, mooddisorder (including depression (e.g., postpartum depression), bipolardisorder, dysthymic disorder, suicidality), schizophrenia or otherpsychotic disorder (including schizoaffective disorder), sleep disorder(including insomnia), substance-related disorder, personality disorder(including obsessive-compulsive personality disorder), autism spectrumdisorders (including those involving mutations to the Shank group ofproteins (e.g., Shank3)), neurodevelopmental disorder (including Rettsyndrome, Tuberous Sclerosis complex), multiple sclerosis, sterolsynthesis disorders, pain (including acute and chronic pain; headaches,e.g., migraine headaches), encephalopathy secondary to a medicalcondition (including hepatic encephalopathy and anti-NMDA receptorencephalitis), acute liver failure, glycine encephalopathy, seizuredisorder (including status epilepticus and monogenic forms of epilepsysuch as Dravet's disease), stroke, traumatic brain injury, movementdisorder (including Huntington's disease and Parkinson's disease),vision impairment, hearing loss, or tinnitus.

In some embodiments, the disorder is Huntington's disease. In someembodiments, the disorder is Parkinson's disease. In some embodiments,the disorder is an inflammatory disease (e.g., lupus).

In some embodiments, the disorder is a sterol synthesis disorder.

In some embodiments, the disorder is Smith-Lemli-Opitz Syndrome (SLOS).In some embodiments, the disorder is desmosterolosis. In someembodiments, the disorder is sitosterolemia. In some embodiments, thedisorder is cerebrotendinous xanthomatosis (CTX). In some embodiments,the disorder is Mevalonate Kinase Deficiency (MKD). In some embodiments,the disorder is SC4MOL gene mutation (SMO Deficiency). In someembodiments, the disorder is Niemann-Pick disease. In some embodiments,the disorder is autism spectrum disorder (ASD). In some embodiments, thedisorder is associated with phenylketomuria.

In an aspect, provided herein is a method of effecting negativeallosteric modulation of an NMDA receptor in a subject, comprisingadministering to the subject a compound described herein, e.g., acompound of Formula (A), a compound of Formula (I-63), or a compound ofFormula (I-67).

Definitions Chemical Definitions

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Isomers can be isolated from mixtures by methods known to those skilledin the art, including chiral high pressure liquid chromatography (HPLC)and the formation and crystallization of chiral salts; or preferredisomers can be prepared by asymmetric syntheses. See, for example,Jacques et al., Enantiomers, Racemates and Resolutions (WileyInterscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977);Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, N Y, 1962); andWilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E. L.Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972). Theinvention additionally encompasses compounds described herein asindividual isomers substantially free of other isomers, andalternatively, as mixtures of various isomers.

The “enantiomeric excess” (“e.e.”) or “% enantiomeric excess” (“% e.e.”)of a composition as used herein refers to an excess of one enantiomerrelative to the other enantiomer present in the composition. Forexample, a composition can contain 90% of one enantiomer, e.g., the Senantiomer, and 10% of the other enantiomer, i.e., the R enantiomer.e.e.=(90−10)/100=80%.

Thus, a composition containing 90% of one enantiomer and 10% of theother enantiomer is said to have an enantiomeric excess of 80%.

The “diastereomeric excess” (“d.e.”) or “% diastereomeric excess” (“%d.e.”) of a composition as used herein refers to an excess of onediastereomer relative to one or more different diasteromers present inthe composition. For example, a composition can contain 90% of onediastereomer, and 10% of one or more different diastereomers.d.e.=(90−10)/100=80%.

Thus, a composition containing 90% of one diastereomers and 10% of oneor more different diastereomers is said to have a diastereomeric excessof 80%.

The absolute configuration of an asymmetric center can be determinedusing methods known to one skilled in the art. In some embodiments, theabsolute configuration of an asymmetric center in a compound can beelucidated from the X-ray single-crystal structure of the compound. Insome embodiments, the absolute configuration of an asymmetric centerelucidated by the X-ray crystal structure of a compound can be used toinfer the absolute configuration of a corresponding asymmetric center inanother compound obtained from the same or similar syntheticmethodologies. In some embodiments, the absolute configuration of anasymmetric center elucidated by the X-ray crystal structure of acompound can be used to infer the absolute configuration of acorresponding asymmetric center in another compound coupled with aspectroscopic technique, e.g., NMR spectroscopy, e.g., ¹H NMRspectroscopy or ¹⁹F NMR spectroscopy.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

The following terms are intended to have the meanings presentedtherewith below and are useful in understanding the description andintended scope of the present invention. When describing the invention,which may include compounds, pharmaceutical compositions containing suchcompounds and methods of using such compounds and compositions, thefollowing terms, if present, have the following meanings unlessotherwise indicated. It should also be understood that when describedherein any of the moieties defined forth below may be substituted with avariety of substituents, and that the respective definitions areintended to include such substituted moieties within their scope as setout below. Unless otherwise stated, the term “substituted” is to bedefined as set out below. It should be further understood that the terms“groups” and “radicals” can be considered interchangeable when usedherein. The articles “a” and “an” may be used herein to refer to one orto more than one (i.e. at least one) of the grammatical objects of thearticle. By way of example “an analogue” means one analogue or more thanone analogue.

“Aliphatic” refers to an alkyl, alkenyl, alkynyl, or carbocyclyl group,as defined herein.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 12 carbon atoms (“C₁₋₁₂alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms(“C₁-C₁₀ alkyl”). In some embodiments, an alkyl group has 1 to 9 carbonatoms (“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8carbon atoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1to 7 carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl grouphas 1 to 6 carbon atoms (“C₁₋₆ alkyl”, also referred to herein as “loweralkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms(“C₁₋₅ alkyl”). In some embodiments, an alkyl group has 1 to 4 carbonatoms (“C₁₋₄ alkyl”). In some embodiments, an alkyl group has 1 to 3carbon atoms (“C₁₋₃ alkyl”). In some embodiments, an alkyl group has 1to 2 carbon atoms (“C₁₋₂ alkyl”). In some embodiments, an alkyl grouphas 1 carbon atom (“C₁ alkyl”). In some embodiments, an alkyl group has2 to 6 carbon atoms (“C₂₋₆ alkyl”). Examples of C₁₋₆ alkyl groupsinclude methyl (C₁), ethyl (C₂), n-propyl (C₃), isopropyl (C₃), n-butyl(C₄), tert-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), n-pentyl (C₅),3-pentanyl (C₅), amyl (C₅), isopentyl (C₅), neopentyl (C₅),3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₅)and the like. Unless otherwise specified, each instance of an alkylgroup is independently optionally substituted, i.e., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents; e.g., for instance from 1 to 5 substituents, 1 to3 substituents, or 1 substituent. In certain embodiments, the alkylgroup is unsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments,the alkyl group is substituted C₁₋₁₀ alkyl. Common alkyl abbreviationsinclude Me (—CH₃), Et (—CH₂CH₃), iPr (—CH(CH₃)₂), nPr (—CH₂CH₂CH₃), n-Bu(—CH₂CH₂CH₂CH₃), or i-Bu (—CH₂CH(CH₃)₂).

“Alkylene” refers to an alkyl group wherein two hydrogens are removed toprovide a divalent radical, and which may be substituted orunsubstituted. Unsubstituted alkylene groups include, but are notlimited to, methylene (—CH₂—), ethylene (—CH₂CH₂—), propylene(—CH₂CH₂CH₂—), butylene (—CH₂CH₂CH₂CH₂—), pentylene (—CH₂CH₂CH₂CH₂CH₂—),hexylene (—CH₂CH₂CH₂CH₂CH₂CH₂—), and the like. Exemplary substitutedalkylene groups, e.g., substituted with one or more alkyl (methyl)groups, include but are not limited to, substituted methylene(—CH(CH₃)—, (—C(CH₃)₂—), substituted ethylene (—CH(CH₃)CH₂—,—CH₂CH(CH₃)—, —C(CH₃)₂CH₂—, —CH₂C(CH₃)₂—), substituted propylene(—CH(CH₃)CH₂CH₂—, —CH₂CH(CH₃)CH₂—, —CH₂CH₂CH(CH₃)—, —C(CH₃)₂CH₂CH₂—,—CH₂C(CH₃)₂CH₂—, —CH₂CH₂C(CH₃)₂—), and the like. When a range or numberof carbons is provided for a particular alkylene group, it is understoodthat the range or number refers to the range or number of carbons in thelinear carbon divalent chain. Alkylene groups may be substituted orunsubstituted with one or more substituents as described herein.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon double bonds (e.g., 1, 2, 3, or 4 carbon-carbon doublebonds), and optionally one or more carbon-carbon triple bonds (e.g., 1,2, 3, or 4 carbon-carbon triple bonds) (“C₂₋₂₀ alkenyl”). In certainembodiments, alkenyl does not contain any triple bonds. In someembodiments, an alkenyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms(“C₂₋₉ alkenyl”). In some embodiments, an alkenyl group has 2 to 8carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, an alkenyl group has2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In some embodiments, an alkenylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”). In some embodiments, analkenyl group has 2 to 5 carbon atoms (“C₂₋₅ alkenyl”). In someembodiments, an alkenyl group has 2 to 4 carbon atoms (“C₂₋₄ alkenyl”).In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C₂₋₃alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C₂alkenyl”). The one or more carbon-carbon double bonds can be internal(such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples ofC₂₋₄ alkenyl groups include ethenyl (C₂), 1-propenyl (C₃), 2-propenyl(C₃), 1-butenyl (C₄), 2-butenyl (C₄), butadienyl (C₄), and the like.Examples of C₂₋₆ alkenyl groups include the aforementioned C₂₋₄ alkenylgroups as well as pentenyl (C₅), pentadienyl (C₅), hexenyl (C₆), and thelike. Additional examples of alkenyl include heptenyl (C₇), octenyl(C₅), octatrienyl (C₅), and the like. Unless otherwise specified, eachinstance of an alkenyl group is independently optionally substituted,i.e., unsubstituted (an “unsubstituted alkenyl”) or substituted (a“substituted alkenyl”) with one or more substituents e.g., for instancefrom 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Incertain embodiments, the alkenyl group is unsubstituted C₂₋₁₀ alkenyl.In certain embodiments, the alkenyl group is substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, one or morecarbon-carbon triple bonds (e.g., 1, 2, 3, or 4 carbon-carbon triplebonds), and optionally one or more carbon-carbon double bonds (e.g., 1,2, 3, or 4 carbon-carbon double bonds) (“C₂₋₂₀ alkynyl”). In certainembodiments, alkynyl does not contain any double bonds. In someembodiments, an alkynyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkynyl”). In some embodiments, an alkynyl group has 2 to 9 carbon atoms(“C₂₋₉ alkynyl”). In some embodiments, an alkynyl group has 2 to 8carbon atoms (“C₂₋₈ alkynyl”). In some embodiments, an alkynyl group has2 to 7 carbon atoms (“C₂₋₇ alkynyl”). In some embodiments, an alkynylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkynyl”). In some embodiments, analkynyl group has 2 to 5 carbon atoms (“C₂₋₅ alkynyl”). In someembodiments, an alkynyl group has 2 to 4 carbon atoms (“C₂₋₄ alkynyl”).In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C₂₋₃alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C₂alkynyl”). The one or more carbon-carbon triple bonds can be internal(such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples ofC₂₋₄ alkynyl groups include, without limitation, ethynyl (C₂),1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl (C₄), 2-butynyl (C₄), andthe like. Examples of C₂₋₆ alkenyl groups include the aforementionedC₂₋₄ alkynyl groups as well as pentynyl (C₅), hexynyl (C₆), and thelike. Additional examples of alkynyl include heptynyl (C₇), octynyl(C₅), and the like. Unless otherwise specified, each instance of analkynyl group is independently optionally substituted, i.e.,unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents; e.g., for instancefrom 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Incertain embodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl.In certain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

The term “heteroalkyl,” as used herein, refers to an alkyl group, asdefined herein, which further comprises 1 or more (e.g., 1, 2, 3, or 4)heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus)within the parent chain, wherein the one or more heteroatoms is insertedbetween adjacent carbon atoms within the parent carbon chain and/or oneor more heteroatoms is inserted between a carbon atom and the parentmolecule, i.e., between the point of attachment. In certain embodiments,a heteroalkyl group refers to a saturated group having from 1 to 10carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC₁₋₁₀ alkyl”). Insome embodiments, a heteroalkyl group is a saturated group having 1 to 9carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC₁₋₉ alkyl”). In someembodiments, a heteroalkyl group is a saturated group having 1 to 8carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC₁₋₈ alkyl”). In someembodiments, a heteroalkyl group is a saturated group having 1 to 7carbon atoms and 1, 2, 3, or 4 heteroatoms (“heteroC₁₋₇ alkyl”). In someembodiments, a heteroalkyl group is a group having 1 to 6 carbon atomsand 1, 2, or 3 heteroatoms (“heteroC₁₋₆ alkyl”). In some embodiments, aheteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1or 2 heteroatoms (“heteroC₁₋₅ alkyl”). In some embodiments, aheteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1or 2 heteroatoms (“heteroC₁₋₄ alkyl”). In some embodiments, aheteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1heteroatom (“heteroC₁₋₃ alkyl”). In some embodiments, a heteroalkylgroup is a saturated group having 1 to 2 carbon atoms and 1 heteroatom(“heteroC₁₋₂ alkyl”). In some embodiments, a heteroalkyl group is asaturated group having 1 carbon atom and 1 heteroatom (“heteroC₁alkyl”). In some embodiments, a heteroalkyl group is a saturated grouphaving 2 to 6 carbon atoms and 1 or 2 heteroatoms (“heteroC₂₋₆ alkyl”).Unless otherwise specified, each instance of a heteroalkyl group isindependently unsubstituted (an “unsubstituted heteroalkyl”) orsubstituted (a “substituted heteroalkyl”) with one or more substituents.In certain embodiments, the heteroalkyl group is an unsubstitutedheteroC₁₋₁₀ alkyl. In certain embodiments, the heteroalkyl group is asubstituted heteroC₁₋₁₀ alkyl.

“‘Aryl” refers to a radical of a monocyclic or polycyclic (e.g.,bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or14 π electrons shared in a cyclic array) having 6-14 ring carbon atomsand zero heteroatoms provided in the aromatic ring system (“C₆₋₁₄aryl”). In some embodiments, an aryl group has six ring carbon atoms(“C₆ aryl”; e.g., phenyl). In some embodiments, an aryl group has tenring carbon atoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and2-naphthyl). In some embodiments, an aryl group has fourteen ring carbonatoms (“C₁₄ aryl”; e.g., anthracyl). “Aryl” also includes ring systemswherein the aryl ring, as defined above, is fused with one or morecarbocyclyl or heterocyclyl groups wherein the radical or point ofattachment is on the aryl ring, and in such instances, the number ofcarbon atoms continue to designate the number of carbon atoms in thearyl ring system. Typical aryl groups include, but are not limited to,groups derived from aceanthrylene, acenaphthylene, acephenanthrylene,anthracene, azulene, benzene, chrysene, coronene, fluoranthene,fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene,indane, indene, naphthalene, octacene, octaphene, octalene, ovalene,penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene,phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene,triphenylene, and trinaphthalene. Particularly aryl groups includephenyl, naphthyl, indenyl, and tetrahydronaphthyl. Unless otherwisespecified, each instance of an aryl group is independently optionallysubstituted, i.e., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

In certain embodiments, an aryl group substituted with one or more ofgroups selected from halo, C₁-C₈ alkyl, C₁-C₈ haloalkyl, cyano, hydroxy,C₁-C₈ alkoxy, and amino.

Examples of representative substituted aryls include the following

wherein one of R⁵⁶ and R⁵⁷ may be hydrogen and at least one of R⁵⁶ andR⁵⁷ is each independently selected from C₁-C₈ alkyl, C₁-C₈ haloalkyl,4-10 membered heterocyclyl, alkanoyl, C₁-C₈ alkoxy, heteroaryloxy,alkylamino, arylamino, heteroarylamino, NR⁵⁸COR⁵⁹, NR⁵⁸SOR⁵⁹ NR⁵⁸SO₂R⁵⁹,COOalkyl, COOaryl, CONR⁵⁸R⁵⁹, CONR⁵⁸OR⁵⁹, NR⁵⁸R⁵⁹, SO₂NR⁵⁸R⁵⁹, S-alkyl,SOalkyl, SO₂alkyl, Saryl, SOaryl, SO₂aryl; or R⁵⁶ and R⁵⁷ may be joinedto form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms,optionally containing one or more heteroatoms selected from the group N,O, or S. R⁶⁰ and R⁶¹ are independently hydrogen, C₁-C₈ alkyl, C₁-C₄haloalkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocyclyl, C₆-C₁₀ aryl,substituted C₆-C₁₀ aryl, 5-10 membered heteroaryl, or substituted 5-10membered heteroaryl.

“Fused aryl” refers to an aryl having two of its ring carbon in commonwith a second aryl or heteroaryl ring or with a carbocyclyl orheterocyclyl ring.

“Heteroaryl” refers to a radical of a 5-10 membered monocyclic orbicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electronsshared in a cyclic array) having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, i.e., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsselected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen,oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unlessotherwise specified, each instance of a heteroaryl group isindependently optionally substituted, i.e., unsubstituted (an“unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”)with one or more substituents. In certain embodiments, the heteroarylgroup is unsubstituted 5-14 membered heteroaryl. In certain embodiments,the heteroaryl group is substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groupsinclude, without limitation, indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

Examples of representative heteroaryls include the following:

wherein each Z is selected from carbonyl, N, NR⁶⁵, O, and S; and R⁶⁵ isindependently hydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 memberedheterocyclyl, C₆-C₁₀ aryl, and 5-10 membered heteroaryl.

“Carbocyclyl” or “carbocyclic” refers to a radical of a non-aromaticcyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C₃₋₁₀carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. Insome embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms(“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to6 ring carbon atoms (“C₃₋₆ carbocyclyl”). In some embodiments, acarbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆ carbocyclyl”). Insome embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms(“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groups include,without limitation, cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl(C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like.Exemplary C₃₋₈ carbocyclyl groups include, without limitation, theaforementioned C₃₋₆ carbocyclyl groups as well as cycloheptyl (C₇),cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇),cyclooctyl (C₈), cyclooctenyl (C₈), bicyclo[2.2.1]heptanyl (C₇),bicyclo[2.2.2]octanyl (C₈), and the like. Exemplary C₃₋₁₀ carbocyclylgroups include, without limitation, the aforementioned C₃₋₈ carbocyclylgroups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀),cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl(C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or contain a fused, bridged orspiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) andcan be saturated or can be partially unsaturated. “Carbocyclyl” alsoincludes ring systems wherein the carbocyclyl ring, as defined above, isfused with one or more aryl or heteroaryl groups wherein the point ofattachment is on the carbocyclyl ring, and in such instances, the numberof carbons continue to designate the number of carbons in thecarbocyclic ring system. Unless otherwise specified, each instance of acarbocyclyl group is independently optionally substituted, i.e.,unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 10 ring carbon atoms (“C₃₋₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). Unless otherwisespecified, each instance of a cycloalkyl group is independentlyunsubstituted (an “unsubstituted cycloalkyl”) or substituted (a“substituted cycloalkyl”) with one or more substituents. In certainembodiments, the cycloalkyl group is unsubstituted C₃₋₁₀ cycloalkyl. Incertain embodiments, the cycloalkyl group is substituted C₃₋₁₀cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to10-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 memberedheterocyclyl”). In heterocyclyl groups that contain one or more nitrogenatoms, the point of attachment can be a carbon or nitrogen atom, asvalency permits. A heterocyclyl group can either be monocyclic(“monocyclic heterocyclyl”) or a fused, bridged or spiro ring systemsuch as a bicyclic system (“bicyclic heterocyclyl”), and can besaturated or can be partially unsaturated. Heterocyclyl bicyclic ringsystems can include one or more heteroatoms in one or both rings.“Heterocyclyl” also includes ring systems wherein the heterocyclyl ring,as defined above, is fused with one or more carbocyclyl groups whereinthe point of attachment is either on the carbocyclyl or heterocyclylring, or ring systems wherein the heterocyclyl ring, as defined above,is fused with one or more aryl or heteroaryl groups, wherein the pointof attachment is on the heterocyclyl ring, and in such instances, thenumber of ring members continue to designate the number of ring membersin the heterocyclyl ring system. Unless otherwise specified, eachinstance of heterocyclyl is independently optionally substituted, i.e.,unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a“substituted heterocyclyl”) with one or more substituents. In certainembodiments, the heterocyclyl group is unsubstituted 3-10 memberedheterocyclyl. In certain embodiments, the heterocyclyl group issubstituted 3-10 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 memberedheterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8membered non-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-6 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-6 membered heterocyclyl”). In some embodiments, the 5-6 memberedheterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen,and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1-2ring heteroatoms selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary4-membered heterocyclyl groups containing one heteroatom include,without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary5-membered heterocyclyl groups containing one heteroatom include,without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyland pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, piperazinyl, morpholinyl,dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containingtwo heteroatoms include, without limitation, triazinanyl. Exemplary7-membered heterocyclyl groups containing one heteroatom include,without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₆ aryl ring (also referred toherein as a 5,6-bicyclic heterocyclic ring) include, without limitation,indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groupsfused to an aryl ring (also referred to herein as a 6,6-bicyclicheterocyclic ring) include, without limitation, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and the like.

“Nitrogen-containing heterocyclyl” group means a 4- to 7-memberednon-aromatic cyclic group containing at least one nitrogen atom, forexample, but without limitation, morpholine, piperidine (e.g.2-piperidinyl, 3-piperidinyl and 4-piperidinyl), pyrrolidine (e.g.2-pyrrolidinyl and 3-pyrrolidinyl), azetidine, pyrrolidone, imidazoline,imidazolidinone, 2-pyrazoline, pyrazolidine, piperazine, and N-alkylpiperazines such as N-methyl piperazine. Particular examples includeazetidine, piperidone and piperazone.

“Hetero” when used to describe a compound or a group present on acompound means that one or more carbon atoms in the compound or grouphave been replaced by a nitrogen, oxygen, or sulfur heteroatom. Heteromay be applied to any of the hydrocarbyl groups described above such asalkyl, e.g., heteroalkyl, cycloalkyl, e.g., heterocyclyl, aryl, e.g,heteroaryl, cycloalkenyl, e.g, cycloheteroalkenyl, and the like havingfrom 1 to 5, and particularly from 1 to 3 heteroatoms.

“Acyl” refers to a radical —C(O)R²⁰, where R²⁰ is hydrogen, substitutedor unsubstituted alkyl, substituted or unsubstituted alkenyl,substituted or unsubstituted alkynyl, substituted or unsubstitutedcarbocyclyl, substituted or unsubstituted heterocyclyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl, asdefined herein. “Alkanoyl” is an acyl group wherein R²⁰ is a group otherthan hydrogen. Representative acyl groups include, but are not limitedto, formyl (—CHO), acetyl (—C(═O)CH₃), cyclohexylcarbonyl,cyclohexylmethylcarbonyl, benzoyl (—C(═O)Ph), benzylcarbonyl(—C(═O)CH₂Ph), —C(O)—C₁-C₈ alkyl, —C(O)—(CH₂)_(t)(C₆-C₁₀ aryl),—C(O)—(CH₂)_(t)(5-10 membered heteroaryl), —C(O)—(CH₂)_(t)(C₃-C₁₀cycloalkyl), and —C(O)—(CH₂)_(t)(4-10 membered heterocyclyl), wherein tis an integer from 0 to 4. In certain embodiments, R²¹ is C₁-C₈ alkyl,substituted with halo or hydroxy; or C₃-C₁₀ cycloalkyl, 4-10 memberedheterocyclyl, C₆-C₁₀ aryl, arylalkyl, 5-10 membered heteroaryl orheteroarylalkyl, each of which is substituted with unsubstituted C₁-C₄alkyl, halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy orhydroxy.

“Alkoxy” refers to the group —OR²⁹ where R²⁹ is substituted orunsubstituted alkyl, substituted or unsubstituted alkenyl, substitutedor unsubstituted alkynyl, substituted or unsubstituted carbocyclyl,substituted or unsubstituted heterocyclyl, substituted or unsubstitutedaryl, or substituted or unsubstituted heteroaryl. Particular alkoxygroups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy.Particular alkoxy groups are lower alkoxy, i.e. with between 1 and 6carbon atoms. Further particular alkoxy groups have between 1 and 4carbon atoms.

In certain embodiments, R²⁹ is a group that has 1 or more substituents,for instance from 1 to 5 substituents, and particularly from 1 to 3substituents, in particular 1 substituent, selected from the groupconsisting of amino, substituted amino, C₆-C₁₀ aryl, aryloxy, carboxyl,cyano, C₃-C₁₀ cycloalkyl, 4-10 membered heterocyclyl, halogen, 5-10membered heteroaryl, hydroxyl, nitro, thioalkoxy, thioaryloxy, thiol,alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—. Exemplary‘substituted alkoxy’ groups include, but are not limited to,—O—(CH₂)_(t)(C₆-C₁₀ aryl), —O—(CH₂)_(t)(5-10 membered heteroaryl),—O—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and —O—(CH₂)_(t)(4-10 memberedheterocyclyl), wherein t is an integer from 0 to 4 and any aryl,heteroaryl, cycloalkyl or heterocyclyl groups present, may themselves besubstituted by unsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄alkoxy, unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl,or unsubstituted C₁-C₄ haloalkoxy or hydroxy. Particular exemplary‘substituted alkoxy’ groups are —OCF₃, —OCH₂CF₃, —OCH₂Ph,—OCH₂-cyclopropyl, —OCH₂CH₂OH, and —OCH₂CH₂NMe₂.

“Amino” refers to the radical —NH₂.

“Oxo group” refers to —C(═O)—.

“Substituted amino” refers to an amino group of the formula —N(R³⁸)₂wherein R³⁸ is hydrogen, substituted or unsubstituted alkyl, substitutedor unsubstituted alkenyl, substituted or unsubstituted alkynyl,substituted or unsubstituted carbocyclyl, substituted or unsubstitutedheterocyclyl, substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, or an amino protecting group, wherein at leastone of R³⁸ is not a hydrogen. In certain embodiments, each R³⁸ isindependently selected from hydrogen, C₁-C₈ alkyl, C₃-C₈ alkenyl, C₃-C₈alkynyl, C₆-C₁₀ aryl, 5-10 membered heteroaryl, 4-10 memberedheterocyclyl, or C₃-C₁₀ cycloalkyl; or C₁-C₈ alkyl, substituted withhalo or hydroxy; C₃-C₈ alkenyl, substituted with halo or hydroxy; C₃-C₈alkynyl, substituted with halo or hydroxy, or —(CH₂)_(t)(C₆-C₁₀ aryl),—(CH₂)_(t)(5-10 membered heteroaryl), —(CH₂)_(t)(C₃-C₁₀ cycloalkyl), or—(CH₂)_(t)(4-10 membered heterocyclyl), wherein t is an integer between0 and 8, each of which is substituted by unsubstituted C₁-C₄ alkyl,halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy orhydroxy; or both R³⁸ groups are joined to form an alkylene group.

Exemplary “substituted amino” groups include, but are not limited to,—NR³⁹—C₁-C₈ alkyl, —NR³⁹—(CH₂)_(t)(C₆-C₁₀ aryl), —NR³⁹—(CH₂)_(t)(5-10membered heteroaryl), —NR³⁹—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—NR³⁹—(CH₂)_(t)(4-10 membered heterocyclyl), wherein t is an integerfrom 0 to 4, for instance 1 or 2, each R³⁹ independently represents H orC₁-C₈ alkyl; and any alkyl groups present, may themselves be substitutedby halo, substituted or unsubstituted amino, or hydroxy; and any aryl,heteroaryl, cycloalkyl, or heterocyclyl groups present, may themselvesbe substituted by unsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄alkoxy, unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl,or unsubstituted C₁-C₄ haloalkoxy or hydroxy. For the avoidance of doubtthe term ‘substituted amino’ includes the groups alkylamino, substitutedalkylamino, alkylarylamino, substituted alkylarylamino, arylamino,substituted arylamino, dialkylamino, and substituted dialkylamino asdefined below. Substituted amino encompasses both monosubstituted aminoand disubstituted amino groups.

“Carboxy” refers to the radical —C(O)OH.

“Cyano” refers to the radical —CN.

“Halo” or “halogen” refers to fluoro (F), chloro (Cl), bromo (Br), andiodo (I). In certain embodiments, the halo group is either fluoro orchloro.

“Haloalkyl” refers to an alkyl radical in which the alkyl group issubstituted with one or more halogens. Typical haloalkyl groups include,but are not limited to, trifluoromethyl (—CF₃), difluoromethyl (—CHF₂),fluoromethyl (—CH₂F), chloromethyl (—CH₂Cl), dichloromethyl (—CHCl₂),tribromomethyl (—CH₂Br), and the like.

“Hydroxy” refers to the radical —OH.

“Nitro” refers to the radical —NO₂.

“Thioketo” refers to the group ═S.

“Carbocyclylalkyl” refers to an alkyl radical in which the alkyl groupis substituted with a cycloalkyl group. Typical carbocyclylalkyl groupsinclude, but are not limited to, cyclopropylmethyl, cyclobutylmethyl,cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl,cyclooctylmethyl, cyclopropylethyl, cyclobutylethyl, cyclopentylethyl,cyclohexylethyl, cycloheptylethyl, and cyclooctylethyl, and the like.

“Heterocyclylalkyl” refers to an alkyl radical in which the alkyl groupis substituted with a heterocyclyl group. Typical heterocyclylalkylgroups include, but are not limited to, pyrrolidinylmethyl,piperidinylmethyl, piperazinylmethyl, morpholinylmethyl,pyrrolidinylethyl, piperidinylethyl, piperazinylethyl, morpholinylethyl,and the like.

“Aralkyl” is a subset of alkyl and aryl, as defined herein, and refersto an optionally substituted alkyl group substituted by an optionallysubstituted aryl group.

Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylgroups, as defined herein, are optionally substituted (e.g.,“substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted”alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or“unsubstituted” carbocyclyl, “substituted” or “unsubstituted”heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or“unsubstituted” heteroaryl group). In general, the term “substituted”,whether preceded by the term “optionally” or not, means that at leastone hydrogen present on a group (e.g., a carbon or nitrogen atom) isreplaced with a permissible substituent, e.g., a substituent which uponsubstitution results in a stable compound, e.g., a compound which doesnot spontaneously undergo transformation such as by rearrangement,cyclization, elimination, or other reaction. Unless otherwise indicated,a “substituted” group has a substituent at one or more substitutablepositions of the group, and when more than one position in any givenstructure is substituted, the substituent is either the same ordifferent at each position. The term “substituted” is contemplated toinclude substitution with all permissible substituents of organiccompounds, any of the substituents described herein that results in theformation of a stable compound. The present invention contemplates anyand all such combinations in order to arrive at a stable compound. Forpurposes of this invention, heteroatoms such as nitrogen may havehydrogen substituents and/or any suitable substituent as describedherein which satisfy the valencies of the heteroatoms and results in theformation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), —NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR^(aa), —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃, —OSi(R^(aa))₃,—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa), —SC(═S)SR^(aa),—SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa), —SC(═O)R^(aa),—P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂, —OP(═O)(R^(aa))₂,—OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, —OP(═O)₂N(R^(bb))₂,—P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂, —NR^(bb)P(═O)(OR^(cc))₂,—NR^(bb)P(═O)(NR^(bb))₂, —P(R^(cc))₂, —P(R^(cc))₃, —OP(R^(cc))₂,—OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂, —BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl,C₁₋₁₀ haloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl, 3-14membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd)groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ haloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl, 3-14membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, or twoR^(aa) groups are joined to form a 3-14 membered heterocyclyl or 5-14membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ haloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ haloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(cc) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R)₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,—NR^(ff)SO₂R^(ee), —SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, 3-10 memberedheterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups, or twogeminal R^(dd) substituents can be joined to form ═O or ═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl,3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, ortwo R^(ff) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃-C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X⁻ is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, SO₄ ⁻²sulfonateions (e.g., methansulfonate, trifluoromethanesulfonate,p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate,naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate,ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate ions(e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate,tartrate, glycolate, and the like).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quaternary nitrogen atoms.Exemplary nitrogen atom substitutents include, but are not limited to,hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R^(aa),—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ haloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The invention is notintended to be limited in any manner by the above exemplary listing ofsubstituents.

Other Definitions

The term “pharmaceutically acceptable salt” refers to those salts whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, Berge et al.,describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptablesalts of the compounds of this invention include those derived fromsuitable inorganic and organic acids and bases. Examples ofpharmaceutically acceptable, nontoxic acid addition salts are salts ofan amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, oxalic acid, maleic acid,tartaric acid, citric acid, succinic acid or malonic acid or by usingother methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Pharmaceutically acceptable salts derived from appropriatebases include alkali metal, alkaline earth metal, ammonium andN⁺(C₁₋₄alkyl)₄ salts. Representative alkali or alkaline earth metalsalts include sodium, lithium, potassium, calcium, magnesium, and thelike. Further pharmaceutically acceptable salts include, whenappropriate, nontoxic ammonium, quaternary ammonium, and amine cationsformed using counterions such as halide, hydroxide, carboxylate,sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (i.e., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or anon-human animal, e.g., a mammal such as primates (e.g., cynomolgusmonkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents,cats, and/or dogs. In certain embodiments, the subject is a human. Incertain embodiments, the subject is a non-human animal. The terms“human,” “patient,” and “subject” are used interchangeably herein.

Disease, disorder, and condition are used interchangeably herein.

As used herein, and unless otherwise specified, the terms “treat,”“treating” and “treatment” contemplate an action that occurs while asubject is suffering from the specified disease, disorder or condition,which reduces the severity of the disease, disorder or condition, orretards or slows the progression of the disease, disorder or condition(“therapeutic treatment”), and also contemplates an action that occursbefore a subject begins to suffer from the specified disease, disorderor condition (“prophylactic treatment”).

In general, the “effective amount” of a compound refers to an amountsufficient to elicit the desired biological response. As will beappreciated by those of ordinary skill in this art, the effective amountof a compound of the invention may vary depending on such factors as thedesired biological endpoint, the pharmacokinetics of the compound, thedisease being treated, the mode of administration, and the age, health,and condition of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

As used herein, and unless otherwise specified, a “therapeuticallyeffective amount” of a compound is an amount sufficient to provide atherapeutic benefit in the treatment of a disease, disorder orcondition, or to delay or minimize one or more symptoms associated withthe disease, disorder or condition. A therapeutically effective amountof a compound means an amount of therapeutic agent, alone or incombination with other therapies, which provides a therapeutic benefitin the treatment of the disease, disorder or condition. The term“therapeutically effective amount” can encompass an amount that improvesoverall therapy, reduces or avoids symptoms or causes of disease orcondition, or enhances the therapeutic efficacy of another therapeuticagent.

As used herein, and unless otherwise specified, a “prophylacticallyeffective amount” of a compound is an amount sufficient to prevent adisease, disorder or condition, or one or more symptoms associated withthe disease, disorder or condition, or prevent its recurrence. Aprophylactically effective amount of a compound means an amount of atherapeutic agent, alone or in combination with other agents, whichprovides a prophylactic benefit in the prevention of the disease,disorder or condition. The term “prophylactically effective amount” canencompass an amount that improves overall prophylaxis or enhances theprophylactic efficacy of another prophylactic agent.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

As generally described herein, the present invention providessubstituted oxysterols useful for preventing and/or treating a broadrange of disorders, including, but not limited to, NMDA-mediateddisorders. These compounds are expected to show improved in vivopotency, pharmacokinetic (PK) properties, oral bioavailability,formulatability, stability, and/or safety as compared to otheroxysterols.

Compounds

In one aspect, provided herein are compounds according to Formula (A):

or a pharmaceutically acceptable salt thereof, wherein:

R¹ is hydrogen or alkyl (e.g., C₁-C₆ alkyl); each of R² and R³ isindependently hydrogen, alkyl (e.g., C₁-C₆ alkyl), alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R² and R³, togetherwith the carbon atom to which they are attached form a 3-8 memberedring; each of R⁴ and R⁵ is independently hydrogen, halo, or —OR^(C),wherein R^(C) is hydrogen or C₁-C₆ alkyl (e.g., C₁-C₃ alkyl), or R⁴ andR⁵, together with the carbon atom to which they are attached form an oxogroup; R⁶ is absent or hydrogen; R^(G) is hydrogen or alkyl; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl (e.g., C₁-C₆ alkyl). In someembodiments, R¹ is C₁-C₆ alkyl (e.g., —CH₃, —CH₂CH₃, —CH₂OCH₃, or —CF₃).In some embodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments,R¹ is —CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., C₁-C₃ alkyl).

In some embodiments, R² is hydrogen or alkyl (e.g., C₁-C₆ alkyl).

In some embodiments, each of R² and R³ is independently hydrogen oralkyl (e.g., C₁-C₆ alkyl). In some embodiments, each of R² and R³ isindependently hydrogen or C₁-C₆ haloalkyl (e.g., —CF₃). In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.

In some embodiments, R⁴ is —OH or halo (e.g., —F).

In some embodiments, R⁴ and R⁵, together with the carbon atom to whichthey are attached form an oxo group. In some embodiments, R⁴ is hydrogenand R⁵ is halo (e.g., —F). In some embodiments, R⁴ and R⁵ are halo(e.g., —F). In some embodiments, R⁴ and R⁵ are hydrogen.

In some embodiments, R² is aryl or heteroaryl and R³ is hydrogen. Insome embodiments, R² is carbocyclyl or heterocyclyl and R³ is hydrogen.In some embodiments, R² and R³ are hydrogen. In some embodiments, R² andR³, together with the carbon atom to which they are attached form a 3-8membered carbocyclic or heterocyclic ring.

In some embodiments, R⁶ is hydrogen and

represents a single bond.

In some embodiments, R^(G) is hydrogen or —CH₃.

In one aspect, provided herein are compounds according to Formula(I-63):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is alkyl(e.g., C₁-C₆ alkyl); each of R² and R³ is independently hydrogen, alkyl(e.g., C₁-C₆ alkyl), alkenyl (e.g., C₂-C₆ alkenyl), alkynyl (e.g., C₂-C₆alkynyl), carbocyclyl, heterocyclyl, aryl, or heteroaryl, or R² and R³,together with the carbon atom to which they are attached form a 3-8membered ring; each of R⁴ and R⁵ is independently hydrogen, halo, or—OR^(C), wherein R^(C) is hydrogen or C₁-C₆ alkyl (e.g., C₁-C₃ alkyl),or R⁴ and R⁵, together with the carbon atom to which they are attachedform an oxo group; R⁶ is absent or hydrogen; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl (e.g., C₁-C₆ alkyl). In someembodiments, R¹ is C₁-C₆ alkyl (e.g., —CH₃, —CH₂CH₃, —CH₂OCH₃, or —CF₃).In some embodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments,R¹ is —CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., C₁-C₃ alkyl). Insome embodiments, R¹ is unsubstituted alkyl. In some embodiments R¹ is—CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., —CH₃).

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroarylor R² and R³, together with the carbon atom to which they are attachedform a 3-8 membered ring.

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), carbocyclyl, heterocyclyl, aryl, or heteroaryl or R² and R³,together with the carbon atom to which they are attached form a 3-8membered ring.

In some embodiments, R² is hydrogen or alkyl (e.g., C₁-C₆ alkyl).

In some embodiments, each of R² and R³ is independently hydrogen oralkyl (e.g., C₁-C₆ alkyl). In some embodiments, each of R² and R³ isindependently hydrogen or C₁-C₆ haloalkyl (e.g., —CF₃). In someembodiments, each of R² and R³ is independently C₅ alkyl (e.g.,substituted or unsubstituted isopentyl) or hydrogen. In someembodiments, each of R² and R³ is independently isopentyl (e.g.,substituted or unsubstituted isopentyl) or hydrogen. In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.

In some embodiments, R⁴ is —OH or halo (e.g., —F).

In some embodiments, R⁴ and R⁵, together with the carbon atom to whichthey are attached form an oxo group. In some embodiments, R⁴ is hydrogenand R⁵ is halo (e.g., —F). In some embodiments, R⁴ and R⁵ are halo(e.g., —F). In some embodiments, R⁴ and R⁵ are hydrogen.

In some embodiments, R² is aryl or heteroaryl and R³ is hydrogen. Insome embodiments, R² is carbocyclyl or heterocyclyl and R³ is hydrogen.In some embodiments, R² and R³ are hydrogen. In some embodiments, R² isisopentyl (e.g., substituted or unsubstituted isopentyl) and R³ ishydrogen. In some embodiments, R² is —CF₃ or —CH₃ and R³ is hydrogen or—CH₃. In some embodiments, R² and R³, together with the carbon atom towhich they are attached form a 3-8 membered carbocyclic or heterocyclicring.

In some embodiments, R¹ is —CH₃ or —CH₂CH₃, R² is isopentyl (e.g.,substituted or unsubstituted isopentyl), and R³ is hydrogen. In someembodiments, R¹ is —CH₃ or —CH₂CH₃, R² is unsubstituted isopentyl, andR³ is hydrogen.

In some embodiments, R² is unsubstituted C₁-C₆ alkyl or C₁-C₆ haloalkyl.In some embodiments, R² is unsubstituted C₁-C₆ alkyl. In someembodiments, R² is pyridyl. In some embodiments, each of R² is isopentyland R³ is hydrogen. In some embodiments, R² is —CF₃ and R³ is hydrogen.In some embodiments, R² is unsubstituted alkyl (e.g., unsubstitutedC₁-C₆ alkyl). In some embodiments, R² is carbocyclylalkyl. In someembodiments, R² is carbocyclylalkyl and R³ is hydrogen. In someembodiments, R² is aralkyl (e.g., benzyl). In some embodiments, R² isheterocyclylalkyl. In some embodiments, wherein R² is unsubstitutedC₁-C₆ alkyl, C₁-C₆ haloalkyl, carbocyclyl, carbocyclylalkyl, aralkyl, orheterocyclylalkyl.

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-A63), (I-B63), or (I-C63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-A63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-B63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-C63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-D63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-E63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-D-i63) or (I-D-ii63):

In some embodiments, the compound of Formula (I-63) is selected from acompound of Formula (I-E-i63) or (I-E-ii63):

In one aspect, provided herein are compounds according to Formula(I-67):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogenor alkyl (e.g., C₁-C₆ alkyl); each of R² and R³ is independentlyhydrogen, alkyl (e.g., C₁-C₆ alkyl), carbocyclyl, heterocyclyl, aryl, orheteroaryl, or R² and R³, together with the carbon atom to which theyare attached form a 3-8 membered ring; each of R⁴ and R⁵ isindependently hydrogen, halo, or —OR^(C), wherein R^(C) is hydrogen orC₁-C₆ alkyl (e.g., C₁-C₃ alkyl), or R⁴ and R⁵, together with the carbonatom to which they are attached form an oxo group; R⁶ is absent orhydrogen; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and when both of

are single bonds, then R⁶ is hydrogen.

In some embodiments, R¹ is alkyl. In some embodiments, R¹ isunsubstituted alkyl. In some embodiments, R¹ is C₁-C₆ alkyl. In someembodiments, R¹ is —CH₃, —CF₃, or —CH₂CH₃. In some embodiments, R¹ is—CH₂OR^(A), wherein R^(A) is C₁-C₆ alkyl (e.g., —CH₃).

In some embodiments, R² is alkyl (e.g., C₁-C₆ alkyl), carbocyclyl,heterocyclyl, aryl, or heteroaryl and R³ is hydrogen, alkyl (e.g., C₁-C₆alkyl), carbocyclyl, heterocyclyl, aryl, or heteroaryl or R² and R³,together with the carbon atom to which they are attached form a 3-8membered ring.

In some embodiments, R² is hydrogen or alkyl. In some embodiments, eachof R² and R³ is independently hydrogen or alkyl. In some embodiments,each of R² and R³ is independently hydrogen or C₁-C₆ haloalkyl. In someembodiments, each of R² and R³ is independently hydrogen, —CF₃, or —CH₃.In some embodiments, R⁴ is —OH or halo. In some embodiments, R⁴ and R⁵,together with the carbon atom to which they are attached form an oxogroup. In some embodiments, R⁴ is hydrogen and R⁵ is halo. In someembodiments, R⁴ and R⁵ are halo. In some embodiments, R⁴ and R⁵ arehydrogen. In some embodiments, R² is aryl or heteroaryl and R³ ishydrogen. In some embodiments, R² is carbocyclyl or heterocyclyl and R³is hydrogen. In some embodiments, R² and R³ are hydrogen. In someembodiments, R² and R³, together with the carbon atom to which they areattached form a 3-8 membered carbocyclic or heterocyclic ring.

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-A67), (I-B67), or (I-C67):

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-A67):

In some embodiments, the compound of Formula (I-67) is selected from acompound of Formula (I-C67):

Exemplary compounds of the invention include:

Alternative Embodiments

In an alternative embodiment, compounds described herein may alsocomprise one or more isotopic substitutions. For example, hydrogen maybe ²H (D or deuterium) or ³H (T or tritium); carbon may be, for example,¹³C or ¹⁴C; oxygen may be, for example, ¹⁸O; nitrogen may be, forexample, ¹⁵N, and the like. In other embodiments, a particular isotope(e.g., ³H, ¹³C, ¹⁴C, ¹⁸O, or ¹⁵N) can represent at least 1%, at least5%, at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, at least 99%, or at least 99.9% of thetotal isotopic abundance of an element that occupies a specific site ofthe compound.

Pharmaceutical Compositions

In another aspect, the invention provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and an effective amountof a compound described herein, or a pharmaceutically acceptable saltthereof.

When employed as pharmaceuticals, the compounds provided herein aretypically administered in the form of a pharmaceutical composition. Suchcompositions can be prepared in a manner well known in thepharmaceutical art and comprise at least one active compound.

In one embodiment, with respect to the pharmaceutical composition, thecarrier is a parenteral carrier, oral or topical carrier.

The present invention also relates to a compound described herein orpharmaceutical composition thereof for use as a pharmaceutical or amedicament.

Generally, the compounds provided herein are administered in atherapeutically effective amount. The amount of the compound actuallyadministered will typically be determined by a physician, in the lightof the relevant circumstances, including the condition to be treated,the chosen route of administration, the actual compound administered,the age, weight, and response of the individual patient, the severity ofthe patient's symptoms, and the like.

The pharmaceutical compositions provided herein can be administered by avariety of routes including oral, rectal, transdermal, subcutaneous,intravenous, intramuscular, and intranasal. Depending on the intendedroute of delivery, the compounds provided herein are preferablyformulated as either injectable or oral compositions or as salves, aslotions or as patches all for transdermal administration.

The compositions for oral administration can take the form of bulkliquid solutions or suspensions, or bulk powders. More commonly,however, the compositions are presented in unit dosage forms tofacilitate accurate dosing. The term “unit dosage forms” refers tophysically discrete units suitable as unitary dosages for human subjectsand other mammals, each unit containing a predetermined quantity ofactive material calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient. Typical unitdosage forms include prefilled, premeasured ampules or syringes of theliquid compositions or pills, tablets, capsules or the like in the caseof solid compositions. In such compositions, the compound is usually aminor component (from about 0.1 to about 50% by weight or preferablyfrom about 1 to about 40% by weight) with the remainder being variousvehicles or carriers and processing aids helpful for forming the desireddosing form.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like. Solid forms may include, forexample, any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art. As before, the active compound in such compositions istypically a minor component, often being from about 0.05 to 10% byweight with the remainder being the injectable carrier and the like.

Transdermal compositions are typically formulated as a topical ointmentor cream containing the active ingredient(s), generally in an amountranging from about 0.01 to about 20% by weight, preferably from about0.1 to about 20% by weight, preferably from about 0.1 to about 10% byweight, and more preferably from about 0.5 to about 15% by weight. Whenformulated as a ointment, the active ingredients will typically becombined with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream with,for example an oil-in-water cream base. Such transdermal formulationsare well-known in the art and generally include additional ingredientsto enhance the dermal penetration of stability of the active ingredientsor the formulation. All such known transdermal formulations andingredients are included within the scope provided herein.

The compounds provided herein can also be administered by a transdermaldevice. Accordingly, transdermal administration can be accomplishedusing a patch either of the reservoir or porous membrane type, or of asolid matrix variety.

The above-described components for orally administrable, injectable ortopically administrable compositions are merely representative. Othermaterials as well as processing techniques and the like are set forth inPart 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, MackPublishing Company, Easton, Pa., which is incorporated herein byreference.

The above-described components for orally administrable, injectable, ortopically administrable compositions are merely representative. Othermaterials as well as processing techniques and the like are set forth inPart 8 of Remington's The Science and Practice of Pharmacy, 21stedition, 2005, Publisher: Lippincott Williams & Wilkins, which isincorporated herein by reference.

The compounds of this invention can also be administered in sustainedrelease forms or from sustained release drug delivery systems. Adescription of representative sustained release materials can be foundin Remington's Pharmaceutical Sciences.

The present invention also relates to the pharmaceutically acceptableformulations of a compound described herein. In one embodiment, theformulation comprises water. In another embodiment, the formulationcomprises a cyclodextrin derivative. The most common cyclodextrins areα-, β- and γ-cyclodextrins consisting of 6, 7 and 8 α-1,4-linked glucoseunits, respectively, optionally comprising one or more substituents onthe linked sugar moieties, which include, but are not limited to,methylated, hydroxyalkylated, acylated, and sulfoalkylethersubstitution. In certain embodiments, the cyclodextrin is a sulfoalkylether β-cyclodextrin, e.g., for example, sulfobutyl etherβ-cyclodextrin, also known as Captisol®. See, e.g., U.S. Pat. No.5,376,645. In certain embodiments, the formulation compriseshexapropyl-β-cyclodextrin. In a more particular embodiment, theformulation comprises hexapropyl-β-cyclodextrin (10-50% in water).

The present invention also relates to the pharmaceutically acceptableacid addition salt of a compound of described herein. The acid which maybe used to prepare the pharmaceutically acceptable salt is that whichforms a non-toxic acid addition salt, i.e., a salt containingpharmacologically acceptable anions such as the hydrochloride,hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate,acetate, lactate, citrate, tartrate, succinate, maleate, fumarate,benzoate, para-toluenesulfonate, and the like.

The following formulation examples illustrate representativepharmaceutical compositions that may be prepared in accordance with thisinvention. The present invention, however, is not limited to thefollowing pharmaceutical compositions.

Exemplary Formulation 1

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 240-270 mg tablets(80-90 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 2

Capsules: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a starch diluent in anapproximate 1:1 weight ratio. The mixture is filled into 250 mg capsules(125 mg of active compound per capsule).

Exemplary Formulation 3

Liquid: A compound described herein, or pharmaceutically acceptable saltthereof, (125 mg) may be admixed with sucrose (1.75 g) and xanthan gum(4 mg) and the resultant mixture may be blended, passed through a No. 10mesh U.S. sieve, and then mixed with a previously made solution ofmicrocrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50mg) in water. Sodium benzoate (10 mg), flavor, and color are dilutedwith water and added with stirring. Sufficient water may then be addedto produce a total volume of 5 mL.

Exemplary Formulation 4

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 450-900 mg tablets(150-300 mg of active compound) in a tablet press.

Exemplary Formulation 5

Injection: A compound described herein, or pharmaceutically acceptablesalt thereof, may be dissolved or suspended in a buffered sterile salineinjectable aqueous medium to a concentration of approximately 5 mg/mL.

Exemplary Formulation 6

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 90-150 mg tablets(30-50 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 7

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be may be admixed as a dry powder with a dry gelatinbinder in an approximate 1:2 weight ratio. A minor amount of magnesiumstearate is added as a lubricant. The mixture is formed into 30-90 mgtablets (10-30 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 8

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 0.3-30 mg tablets(0.1-10 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 9

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 150-240 mg tablets(50-80 mg of active compound per tablet) in a tablet press.

Exemplary Formulation 10

Tablets: A compound described herein, or pharmaceutically acceptablesalt thereof, may be admixed as a dry powder with a dry gelatin binderin an approximate 1:2 weight ratio. A minor amount of magnesium stearateis added as a lubricant. The mixture is formed into 270-450 mg tablets(90-150 mg of active compound per tablet) in a tablet press.

Injection dose levels range from about 0.1 mg/kg/hour to at least 10mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kgor more may also be administered to achieve adequate steady statelevels. The maximum total dose is not expected to exceed about 2 g/dayfor a 40 to 80 kg human patient.

For the prevention and/or treatment of long-term conditions the regimenfor treatment usually stretches over many months or years so oral dosingis preferred for patient convenience and tolerance. With oral dosing,one to five and especially two to four and typically three oral dosesper day are representative regimens. Using these dosing patterns, eachdose provides from about 0.01 to about 20 mg/kg of the compound providedherein, with preferred doses each providing from about 0.1 to about 10mg/kg, and especially about 1 to about 5 mg/kg.

Transdermal doses are generally selected to provide similar or lowerblood levels than are achieved using injection doses.

When used to prevent the onset of a CNS-disorder, the compounds providedherein will be administered to a subject at risk for developing thecondition, typically on the advice and under the supervision of aphysician, at the dosage levels described above. Subjects at risk fordeveloping a particular condition generally include those that have afamily history of the condition, or those who have been identified bygenetic testing or screening to be particularly susceptible todeveloping the condition.

Methods of Treatment and Use

Compounds of the present invention, e.g., a compound of Formula (A),(I-63), or (I-67), and pharmaceutically acceptable salts thereof, asdescribed herein, are generally designed to modulate NMDA function, andtherefore to act as oxysterols for the treatment and prevention of,e.g., CNS-related conditions in a subject. In some embodiments, thecompounds described herein, e.g., a compound of Formula (A), (I-63), or(I-67), and pharmaceutically acceptable salts thereof, as describedherein, are generally designed to penetrate the blood brain barrier(e.g., designed to be transported across the blood brain barrier).Modulation, as used herein, refers to, for example, the inhibition orpotentiation of NMDA receptor function. In certain embodiments, acompound described herein, e.g., a compound of Formula (A), (I-63), or(I-67), or pharmaceutically acceptable salt thereof, acts as a negativeallosteric modulator (NAM) of NMDA receptor function, and inhibit NMDAreceptor function. In certain embodiments, a compound described herein,e.g., a compound of Formula (A), (I-63), or (I-67), or pharmaceuticallyacceptable salt thereof, acts as a positive allosteric modulator (PAM)of NMDA receptor function, and potentiate NMDA receptor function. Incertain embodiments, a compound described herein, e.g., a compound ofFormula (A), (I-63), or (I-67), or pharmaceutically acceptable saltthereof, blocks or reduces the potentiation or inhibition of NMDAreceptor function by a naturally-occurring substrate. Such compounds donot act as negative allosteric modulators (NAMs) or positive allostericmodulators (PAMs) of NMDA receptor function—these compounds can bereferred to as neutral allosteric ligands (NALs) In some embodiments,the disorder is cancer. In some embodiments, the disorder is diabetes.In some embodiments, the disorder is a sterol synthesis disorder. Insome embodiments, the disorder is a gastrointestinal (GI) disorder,e.g., constipation, irritable bowel syndrome (IBS), inflammatory boweldisease (IBD) (e.g., ulcerative colitis, Crohn's disease), structuraldisorders affecting the GI, anal disorders (e.g., hemorrhoids, internalhemorrhoids, external hemorrhoids, anal fissures, perianal abscesses,anal fistula), colon polyps, cancer, or colitis. In some embodiments,the disorder is inflammatory bowel disease.

Exemplary conditions related to NMDA-modulation include, but are notlimited to, gastrointestinal (GI) disorder, e.g., constipation,irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) (e.g.,ulcerative colitis, Crohn's disease), structural disorders affecting theGI, anal disorders (e.g., hemorrhoids, internal hemorrhoids, externalhemorrhoids, anal fissures, perianal abscesses, anal fistula), colonpolyps, cancer, colitis, and CNS conditions, e.g., as described herein.

Exemplary conditions (e.g., CNS conditions) related to NMDA-modulationinclude, but are not limited to, adjustment disorders, anxiety disorders(including obsessive-compulsive disorder, posttraumatic stress disorder,social phobia, generalized anxiety disorder), cognitive disorders(including Alzheimer's disease and other forms of dementia includingcortico-basal dementia-progressive supranucelar palsy, frontal-temporaldementia, primary progressive aphasia, Parkinson's disease dementia, andLewy body dementia), dissociative disorders, eating disorders, mooddisorders (including depression (e.g., postpartum depression), bipolardisorder, dysthymic disorder, suicidality), schizophrenia or otherpsychotic disorders (including schizoaffective disorder), sleepdisorders (including insomnia), substance abuse-related disorders,personality disorders (including obsessive-compulsive personalitydisorder), autism spectrum disorders (including those involvingmutations to the Shank group of proteins (e.g., Shank3)),neurodevelopmental disorders (including Rett syndrome), multiplesclerosis, sterol synthesis disorders, Smith-Lemli-Opitz syndrome, pain(including acute pain, chronic pain, and neuropathic pain), seizuredisorders (including status epilepticus and monogenic forms of epilepsysuch as Dravet's disease, Tuberous Sclerosis Complex (TSC), andinfantile spasms), stroke, subarachnoid hemorrhage, intracerebralhemorrhage, cerebral ischemia, traumatic brain injury, movementdisorders (including Huntington's disease and Parkinson's disease)attention deficit disorder, attention deficit hyperactivity disorder,metabolic encephalopathies (including phenylketoneuria), post-partumpsychosis, syndromes associated with high titers of anti-NMDA receptorantibodies (including anti-NMDA receptor encephalitis),neurodegenerative disorders, neuroinflammation, neuropsychiatric lupus,Niemann-Pick C disorder, and tinnitus.

In certain embodiments, compounds of the present invention, e.g., acompound described herein, e.g., a compound of Formula (A), (I-63), or(I-67), or pharmaceutically acceptable salt thereof, can be used toinduce sedation or anesthesia.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofadjustment disorders, anxiety disorders (including obsessive-compulsivedisorder, posttraumatic stress disorder, social phobia, generalizedanxiety disorder), cognitive disorders (including Alzheimer's diseaseand other forms of dementia including cortico-basal dementia-progressivesupranucelar palsy, frontal-temoral dementia, primary progressiveaphasia, Parkinson's disease dementia, and Lewy body dementia),dissociative disorders, eating disorders, mood disorders (includingdepression (e.g., postpartum depression), bipolar disorder, dysthymicdisorder, suicidality), schizophrenia or other psychotic disorders(including schizoaffective disorder), sleep disorders (includinginsomnia), substance abuse-related disorders, personality disorders(including obsessive-compulsive personality disorder), autism spectrumdisorders (including those involving mutations to the Shank group ofproteins (e.g., Shank3)), neurodevelopmental disorders (including Rettsyndrome), multiple sclerosis, sterol synthesis disorders,Smith-Lemli-Opitz syndrome, pain (including acute pain, chronic pain,and neuropathic pain), seizure disorders (including status epilepticusand monogenic forms of epilepsy such as Dravet's disease, TuberousSclerosis Complex (TSC), and infantile spasms), stroke, subarachnoidhemorrhage, intracerebral hemorrhage, cerebral ischemia, traumatic braininjury, movement disorders (including Huntington's disease andParkinson's disease) attention deficit disorder, attention deficithyperactivity disorder, metabolic encephalopathies (includingphenylketoneuria), post-partum psychosis, syndromes associated with hightiters of anti-NMDA receptor antibodies (including anti-NMDA receptorencephalitis), neurodegenerative disorders, neuroinflammation,neuropsychiatric lupus, Niemann-Pick C disorder, and tinnitus.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofadjustment disorders, anxiety disorders (including obsessive-compulsivedisorder, posttraumatic stress disorder, social phobia, generalizedanxiety disorder), cognitive disorders (including Alzheimer's diseaseand other forms of dementia including cortico-basal dementia-progressivesupranucelar palsy, frontal-temoral dementia, primary progressiveaphasia, Parkinson's disease dementia, and Lewy body dementia),substance abuse-related disorders, dissociative disorders, eatingdisorders mood disorders (including depression (e.g., postpartumdepression), bipolar disorder, dysthymic disorder, suicidality),schizophrenia or other psychotic disorders (including schizoaffectivedisorder), personality disorders (including obsessive-compulsivepersonality disorder), autism spectrum disorders (including thoseinvolving mutations to the Shank group of proteins (e.g., Shank3)), orpost-partum psychosis.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofneurodevelopmental disorders (including Rett syndrome), multiplesclerosis, sterol synthesis disorders, Smith-Lemli-Opitz syndrome, pain(including acute pain, chronic pain, and neuropathic pain), seizuredisorders (including status epilepticus and monogenic forms of epilepsysuch as Dravet's disease, Tuberous Sclerosis Complex (TSC), andinfantile spasms), stroke, subarachnoid hemorrhage, intracerebralhemorrhage, cerebral ischemia, traumatic brain injury, movementdisorders (including Huntington's disease and Parkinson's disease)attention deficit disorder, attention deficit hyperactivity disorder,metabolic encephalopathies (including phenylketoneuria), syndromesassociated with high titers of anti-NMDA receptor antibodies (includinganti-NMDA receptor encephalitis), neurodegenerative disorders,neuroinflammation, neuropsychiatric lupus, Niemann-Pick C disorder, ortinnitus.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofobsessive compulsive disorder, depression, neuropsychiatric lupus, orschizophrenia.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofHuntington's disease, amyotrophic lateral sclerosis, multiple sclerosis,Alzheimer's disease, dementia, Parkinson's disease, ataxia, Fragile Xsyndrome, Tourette syndrome, levodopa-induced dyskinesia, Rett syndrome,autism spectrum disorder, or traumatic brain injury.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention oftinnitus, neuropathic pain, or migraine.

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofacute liver failure or glycine encephalopathy,

In certain embodiments, a compound described herein, or pharmaceuticallyacceptable salt thereof, is useful in the treatment or prevention ofseizures or genetic epilepsy.

In some embodiments, a compound of the invention, a compound describedherein, e.g., a compound of Formula (A), a compound of Formula (I-63),or a compound of Formula (I-67) that acts as a PAM of NMDA receptorfunction can be useful in the treatment or prevention of conditions(e.g., CNS-related conditions) including schizophrenia or otherpsychotic disorders (including schizoaffective disorder), sleepdisorders (including insomnia), autism spectrum disorders (includingthose involving mutations to the Shank group of proteins (e.g.,Shank3)), multiple sclerosis, movement disorders (including Huntington'sdisease and Parkinson's disease), attention deficit disorder, attentiondeficit hyperactivity disorder, metabolic encephalopathies (includingphenylketoneuria), post-partum psychosis, and syndromes associated withhigh titers or anti-NMDA receptor antibodies (including anti-NMDAreceptor encephalitis).

In some embodiments, a compound of the invention, e.g., a compound ofFormula (A), a compound of Formula (I-63), or a compound of Formula(I-67), that acts as a NAM of NMDA receptor function can be useful inthe treatment or prevention of conditions (e.g., CNS-related conditions)including anxiety disorders (including obsessive-compulsive disorder,posttraumatic stress disorder, social phobia, generalized anxietydisorder), mood disorders (including depression (e.g., postpartumdepression), bipolar disorder, dysthymic disorder, suicidality),personality disorders (including obsessive-compulsive personalitydisorder), neurodevelopmental disorders (including Rett syndrome), pain(including acute and chronic pain), seizure disorders (including statusepilepticus and monogenic forms of epilepsy such as Dravet's disease,and Tuberous Sclerosis Complex (TSC)), stroke, traumatic brain injury,adjustment disorders, neuropsychiatric lupus, and tinnitus.

In some embodiments, a compound of the invention, e.g., a compound ofFormula (A), a compound of Formula (I-63), or a compound of Formula(I-67), that acts as a PAM or a NAM of NMDA receptor function can beuseful in the treatment or prevention of conditions (e.g., CNS-relatedconditions) including cognitive disorders (including Alzheimer's diseaseand other forms of dementia including cortico-basal dementia-progressivesupranucelar palsy, frontal-temoral dementia, primary progressiveaphasia, Parkinson's disease dementia, and Lewy body dementia), sterolsynthesis disorders, and eating disorders.

In another aspect, provided is a method of treating or preventing brainexcitability in a subject susceptible to or afflicted with a conditionassociated with brain excitability, comprising administering to thesubject an effective amount of a compound of the present invention,e.g., a compound of Formula (A), a compound of Formula (I-63), or acompound of Formula (I-67), or a pharmaceutically acceptable saltthereof.

In yet another aspect, the present invention provides a combination of acompound of the present invention, e.g., a compound of Formula (A), acompound of Formula (I-63), or a compound of Formula (I-67), orpharmaceutically acceptable salt thereof, and another pharmacologicallyactive agent. The compounds provided herein can be administered as thesole active agent or they can be administered in combination with otheragents. Administration in combination can proceed by any techniqueapparent to those of skill in the art including, for example, separate,sequential, concurrent and alternating administration.

In another aspect, the present invention provides a method of effectingnegative allosteric modulation of an NMDA receptor in a subject,comprising administering to the subject a compound described herein,e.g., a compound of Formula (A), a compound of Formula (I-63), or acompound of Formula (I-67).

Movement Disorders

Also described herein are methods for treating a movement disorder. Asused herein, “movement disorders” refers to a variety of diseases anddisorders that are associated with hyperkinetic movement disorders andrelated abnormalities in muscle control. Exemplary movement disordersinclude, but are not limited to, Parkinson's disease and Parkinsonism(defined particularly by bradykinesia), dystonia, chorea andHuntington's disease, ataxia, levodopa-induced dyskinesia, tremor (e.g.,essential tremor), myoclonus and startle, tics and Tourette syndrome,Restless legs syndrome, stiff person syndrome, and gait disorders.

Tremor is an involuntary, at times rhythmic, muscle contraction andrelaxation that can involve oscillations or twitching of one or morebody parts (e.g., hands, arms, eyes, face, head, vocal folds, trunk,legs). Tremor includes hereditary, degenerative, and idiopathicdisorders such as Wilson's disease, Parkinson's disease, and essentialtremor, respectively; metabolic diseases (e.g., thyroid-parathyroid-,liver disease and hypoglycemia); peripheral neuropathies (associatedwith Charcot-Marie-Tooth, Roussy-Levy, diabetes mellitus, complexregional pain syndrome); toxins (nicotine, mercury, lead, CO, Manganese,arsenic, toluene); drug-induced (narcoleptics, tricyclics, lithium,cocaine, alcohol, adrenaline, bronchodilators, theophylline, caffeine,steroids, valproate, amiodarone, thyroid hormones, vincristine); andpsychogenic disorders. Clinical tremor can be classified intophysiologic tremor, enhanced physiologic tremor, essential tremorsyndromes (including classical essential tremor, primary orthostatictremor, and task- and position-specific tremor), dystonic tremor,parkinsonian tremor, cerebellar tremor, Holmes' tremor (i.e., rubraltremor), palatal tremor, neuropathic tremor, toxic or drug-inducedtremor, and psychogenic tremor. Other forms of tremor include cerebellartremor or intention tremor, dystonic tremor, essential tremor,orthostatic tremor, parkinsonian tremor, physiological tremor,psychogenic tremor, or rubral tremor.

Cerebellar tremor or intention tremor is a slow, broad tremor of theextremities that occurs after a purposeful movement. Cerebellar tremoris caused by lesions in or damage to the cerebellum resulting from,e.g., tumor, stroke, disease (e.g., multiple sclerosis, an inheriteddegenerative disorder).

Dystonic tremor occurs in individuals affected by dystonia, a movementdisorder in which sustained involuntary muscle contractions causetwisting and repetitive motions and/or painful and abnormal postures orpositions. Dystonic tremor may affect any muscle in the body. Dystonictremors occurs irregularly and often can be relieved by complete rest.

Essential tremor or benign essential tremor is the most common type oftremor. Essential tremor may be mild and nonprogressive in some, and maybe slowly progressive, starting on one side of the body but affect bothsides within 3 years. The hands are most often affected, but the head,voice, tongue, legs, and trunk may also be involved. Tremor frequencymay decrease as the person ages, but severity may increase. Heightenedemotion, stress, fever, physical exhaustion, or low blood sugar maytrigger tremors and/or increase their severity. Symptoms generallyevolve over time and can be both visible and persistent following onset.

Orthostatic tremor is characterized by fast (e.g., greater than 12 Hz)rhythmic muscle contractions that occurs in the legs and trunkimmediately after standing. Cramps are felt in the thighs and legs andthe patient may shake uncontrollably when asked to stand in one spot.Orthostatic tremor may occur in patients with essential tremor.

Parkinsonian tremor is caused by damage to structures within the brainthat control movement. Parkinsonian tremor is often a precursor toParkinson's disease and is typically seen as a “pill-rolling” action ofthe hands that may also affect the chin, lips, legs, and trunk. Onset ofparkinsonian tremor typically begins after age 60. Movement starts inone limb or on one side of the body and can progress to include theother side.

Physiological tremor can occur in normal individuals and have noclinical significance. It can be seen in all voluntary muscle groups.Physiological tremor can be caused by certain drugs, alcohol withdrawal,or medical conditions including an overactive thyroid and hypoglycemia.The tremor classically has a frequency of about 10 Hz.

Psychogenic tremor or hysterical tremor can occur at rest or duringpostural or kinetic movement. Patient with psychogenic tremor may have aconversion disorder or another psychiatric disease.

Rubral tremor is characterized by coarse slow tremor which can bepresent at rest, at posture, and with intention. The tremor isassociated with conditions that affect the red nucleus in the midbrain,classical unusual strokes.

Parkinson's disease affects nerve cells in the brain that producedopamine. Symptoms include muscle rigidity, tremors, and changes inspeech and gait. Parkinsonism is characterized by tremor, bradykinesia,rigidity, and postural instability. Parkinsonism shares symptoms foundin Parkinson's disease, but is a symptom complex rather than aprogressive neurodegenerative disease.

Dystonia is a movement disorder characterized by sustained orintermittent muscle contractions causing abnormal, often repetitivemovements or postures. Dystonic movements can be patterned, twisting,and may be tremulous. Dystonia is often initiated or worsened byvoluntary action and associated with overflow muscle activation.

Chorea is a neurological disorder characterized by jerky involuntarymovements typically affecting the shoulders, hips, and face.

Huntington's Disease is an inherited disease that causes nerve cells inthe brain to waste away. Symptoms include uncontrolled movements,clumsiness, and balance problems. Huntington's disease can hinder walk,talk, and swallowing.

Ataxia refers to the loss of full control of bodily movements, and mayaffect the fingers, hands, arms, legs, body, speech, and eye movements.

Myoclonus and Startle is a response to a sudden and unexpected stimulus,which can be acoustic, tactile, visual, or vestibular.

Tics are an involuntary movement usually onset suddenly, brief,repetitive, but non-rhythmical, typically imitating normal behavior andoften occurring out of a background of normal activity. Tics can beclassified as motor or vocal, motor tics associated with movements whilevocal tics associated with sound. Tics can be characterized as simple orcomplex. For example simple motor tics involve only a few musclesrestricted to a specific body part.

Tourette Syndrome is an inherited neuropsychiatric disorder with onsetin childhood, characterized by multiple motor tics and at least onevocal tic.

Restless Legs Syndrome is a neurologic sensorimotor disordercharacterized by an overwhelming urge to move the legs when at rest.

Stiff Person Syndrome is a progressive movement disorder characterizedby involuntary painful spasms and rigidity of muscles, usually involvingthe lower back and legs. Stiff-legged gait with exaggerated lumbarhyperlordosis typically results. Characteristic abnormality on EMGrecordings with continuous motor unit activity of the paraspinal axialmuscles is typically observed. Variants include “stiff-limb syndrome”producing focal stiffness typically affecting distal legs and feet.

Gait disorders refer to an abnormality in the manner or style ofwalking, which results from neuromuscular, arthritic, or other bodychanges. Gait is classified according to the system responsible forabnormal locomotion, and include hemiplegic gait, diplegic gait,neuropathic gait, myopathic gait, parkinsonian gait, choreiform gait,ataxic gait, and sensory gait.

Mood Disorders

Also provided herein are methods for treating a mood disorder, forexample clinical depression, postnatal depression or postpartumdepression, perinatal depression, atypical depression, melancholicdepression, psychotic major depression, cationic depression, seasonalaffective disorder, dysthymia, double depression, depressive personalitydisorder, recurrent brief depression, minor depressive disorder, bipolardisorder or manic depressive disorder, depression caused by chronicmedical conditions, treatment-resistant depression, refractorydepression, suicidality, suicidal ideation, or suicidal behavior.

Clinical depression is also known as major depression, major depressivedisorder (MDD), severe depression, unipolar depression, unipolardisorder, and recurrent depression, and refers to a mental disordercharacterized by pervasive and persistent low mood that is accompaniedby low self-esteem and loss of interest or pleasure in normallyenjoyable activities. Some people with clinical depression have troublesleeping, lose weight, and generally feel agitated and irritable.Clinical depression affects how an individual feels, thinks, and behavesand may lead to a variety of emotional and physical problems.Individuals with clinical depression may have trouble doing day-to-dayactivities and make an individual feel as if life is not worth living.

Postnatal depression (PND) is also referred to as postpartum depression(PPD), and refers to a type of clinical depression that affects womenafter childbirth. Symptoms can include sadness, fatigue, changes insleeping and eating habits, reduced sexual desire, crying episodes,anxiety, and irritability. In some embodiments, the PND is atreatment-resistant depression (e.g., a treatment-resistant depressionas described herein). In some embodiments, the PND is refractorydepression (e.g., a refractory depression as described herein).

In some embodiments, a subject having PND also experienced depression,or a symptom of depression during pregnancy. This depression is referredto herein as) perinatal depression. In an embodiment, a subjectexperiencing perinatal depression is at increased risk of experiencingPND.

Atypical depression (AD) is characterized by mood reactivity (e.g.,paradoxical anhedonia) and positivity, significant weight gain orincreased appetite. Patients suffering from AD also may have excessivesleep or somnolence (hypersomnia), a sensation of limb heaviness, andsignificant social impairment as a consequence of hypersensitivity toperceived interpersonal rejection.

Melancholic depression is characterized by loss of pleasure (anhedonia)in most or all activities, failures to react to pleasurable stimuli,depressed mood more pronounced than that of grief or loss, excessiveweight loss, or excessive guilt.

Psychotic major depression (PMD) or psychotic depression refers to amajor depressive episode, in particular of melancholic nature, where theindividual experiences psychotic symptoms such as delusions andhallucinations.

Catatonic depression refers to major depression involving disturbancesof motor behavior and other symptoms. An individual may become mute andstuporose, and either is immobile or exhibits purposeless or bizarremovements.

Seasonal affective disorder (SAD) refers to a type of seasonaldepression wherein an individual has seasonal patterns of depressiveepisodes coming on in the fall or winter.

Dysthymia refers to a condition related to unipolar depression, wherethe same physical and cognitive problems are evident. They are not assevere and tend to last longer (e.g., at least 2 years).

Double depression refers to fairly depressed mood (dysthymia) that lastsfor at least 2 years and is punctuated by periods of major depression.

Depressive Personality Disorder (DPD) refers to a personality disorderwith depressive features.

Recurrent Brief Depression (RBD) refers to a condition in whichindividuals have depressive episodes about once per month, each episodelasting 2 weeks or less and typically less than 2-3 days.

Minor depressive disorder or minor depression refers to a depression inwhich at least 2 symptoms are present for 2 weeks.

Bipolar disorder or manic depressive disorder causes extreme mood swingsthat include emotional highs (mania or hypomania) and lows (depression).During periods of mania the individual may feel or act abnormally happy,energetic, or irritable. They often make poorly thought out decisionswith little regard to the consequences. The need for sleep is usuallyreduced. During periods of depression there may be crying, poor eyecontact with others, and a negative outlook on life. The risk of suicideamong those with the disorder is high at greater than 6% over 20 years,while self-harm occurs in 30-40%. Other mental health issues such asanxiety disorder and substance use disorder are commonly associated withbipolar disorder.

Depression caused by chronic medical conditions refers to depressioncaused by chronic medical conditions such as cancer or chronic pain,chemotherapy, chronic stress.

Treatment-resistant depression refers to a condition where theindividuals have been treated for depression, but the symptoms do notimprove. For example, antidepressants or psychological counseling(psychotherapy) do not ease depression symptoms for individuals withtreatment-resistant depression. In some cases, individuals withtreatment-resistant depression improve symptoms, but come back.Refractory depression occurs in patients suffering from depression whoare resistant to standard pharmacological treatments, includingtricyclic antidepressants, MAOIs, SSRIs, and double and triple uptakeinhibitors and/or anxiolytic drugs, as well as non-pharmacologicaltreatments (e.g., psychotherapy, electroconvulsive therapy, vagus nervestimulation and/or transcranial magnetic stimulation).

Suicidality, suicidal ideation, suicidal behavior refers to the tendencyof an individual to commit suicide. Suicidal ideation concerns thoughtsabout or an unusual preoccupation with suicide. The range of suicidalideation varies greatly, from e.g., fleeting thoughts to extensivethoughts, detailed planning, role playing, incomplete attempts. Symptomsinclude talking about suicide, getting the means to commit suicide,withdrawing from social contact, being preoccupied with death, feelingtrapped or hopeless about a situation, increasing use of alcohol ordrugs, doing risky or self-destructive things, saying goodbye to peopleas if they won't be seen again.

Symptoms of depression include persistent anxious or sad feelings,feelings of helplessness, hopelessness, pessimism, worthlessness, lowenergy, restlessness, difficulty sleeping, sleeplessness, irritability,fatigue, motor challenges, loss of interest in pleasurable activities orhobbies, loss of concentration, loss of energy, poor self-esteem,absence of positive thoughts or plans, excessive sleeping, overeating,appetite loss, insomnia, self-harm, thoughts of suicide, and suicideattempts. The presence, severity, frequency, and duration of symptomsmay vary on a case to case basis. Symptoms of depression, and relief ofthe same, may be ascertained by a physician or psychologist (e.g., by amental state examination).

Anxiety Disorders

Provided herein are methods for treating anxiety disorders. Anxietydisorder is a blanket term covering several different forms of abnormaland pathological fear and anxiety. Current psychiatric diagnosticcriteria recognize a wide variety of anxiety disorders.

Generalized anxiety disorder is a common chronic disorder characterizedby long-lasting anxiety that is not focused on any one object orsituation. Those suffering from generalized anxiety experiencenon-specific persistent fear and worry and become overly concerned witheveryday matters. Generalized anxiety disorder is the most commonanxiety disorder to affect older adults.

In panic disorder, a person suffers from brief attacks of intense terrorand apprehension, often marked by trembling, shaking, confusion,dizziness, nausea, difficulty breathing. These panic attacks, defined bythe APA as fear or discomfort that abruptly arises and peaks in lessthan ten minutes, can last for several hours and can be triggered bystress, fear, or even exercise; although the specific cause is notalways apparent. In addition to recurrent unexpected panic attacks, adiagnosis of panic disorder also requires that said attacks have chronicconsequences: either worry over the attacks' potential implications,persistent fear of future attacks, or significant changes in behaviorrelated to the attacks. Accordingly, those suffering from panic disorderexperience symptoms even outside of specific panic episodes. Often,normal changes in heartbeat are noticed by a panic sufferer, leadingthem to think something is wrong with their heart or they are about tohave another panic attack. In some cases, a heightened awareness(hypervigilance) of body functioning occurs during panic attacks,wherein any perceived physiological change is interpreted as a possiblelife threatening illness (i.e. extreme hypochondriasis).

Obsessive compulsive disorder is a type of anxiety disorder primarilycharacterized by repetitive obsessions (distressing, persistent, andintrusive thoughts or images) and compulsions (urges to perform specificacts or rituals). The OCD thought pattern may be likened tosuperstitions insofar as it involves a belief in a causativerelationship where, in reality, one does not exist. Often the process isentirely illogical; for example, the compulsion of walking in a certainpattern may be employed to alleviate the obsession of impending harm.And in many cases, the compulsion is entirely inexplicable, simply anurge to complete a ritual triggered by nervousness. In a minority ofcases, sufferers of OCD may only experience obsessions, with no overtcompulsions; a much smaller number of sufferers experience onlycompulsions.

The single largest category of anxiety disorders is that of phobia,which includes all cases in which fear and anxiety is triggered by aspecific stimulus or situation. Sufferers typically anticipateterrifying consequences from encountering the object of their fear,which can be anything from an animal to a location to a bodily fluid.

Post-traumatic stress disorder or PTSD is an anxiety disorder whichresults from a traumatic experience. Post-traumatic stress can resultfrom an extreme situation, such as combat, rape, hostage situations, oreven serious accident. It can also result from long term (chronic)exposure to a severe stressor, for example soldiers who endureindividual battles but cannot cope with continuous combat. Commonsymptoms include flashbacks, avoidant behaviors, and depression.

Epilepsy

Epilepsy is a brain disorder characterized by repeated seizures overtime. Types of epilepsy can include, but are not limited to generalizedepilepsy, e.g., childhood absence epilepsy, juvenile myoclonic epilepsy,epilepsy with grand-mal seizures on awakening, West syndrome,Lennox-Gastaut syndrome, partial epilepsy, e.g., temporal lobe epilepsy,frontal lobe epilepsy, benign focal epilepsy of childhood.

Epileptogenesis

Epileptogenesis is a gradual process by which a normal brain developsepilepsy (a chronic condition in which seizures occur). Epileptogenesisresults from neuronal damage precipitated by the initial insult (e.g.,status epilepticus).

Status Epilepticus (SE)

Status epilepticus (SE) can include, e.g., convulsive statusepilepticus, e.g., early status epilepticus, established statusepilepticus, refractory status epilepticus, super-refractory statusepilepticus; non-convulsive status epilepticus, e.g., generalized statusepilepticus, complex partial status epilepticus; generalized periodicepileptiform discharges; and periodic lateralized epileptiformdischarges. Convulsive status epilepticus is characterized by thepresence of convulsive status epileptic seizures, and can include earlystatus epilepticus, established status epilepticus, refractory statusepilepticus, super-refractory status epilepticus. Early statusepilepticus is treated with a first line therapy. Established statusepilepticus is characterized by status epileptic seizures which persistdespite treatment with a first line therapy, and a second line therapyis administered. Refractory status epilepticus is characterized bystatus epileptic seizures which persist despite treatment with a firstline and a second line therapy, and a general anesthetic is generallyadministered. Super refractory status epilepticus is characterized bystatus epileptic seizures which persist despite treatment with a firstline therapy, a second line therapy, and a general anesthetic for 24hours or more.

Non-convulsive status epilepticus can include, e.g., focalnon-convulsive status epilepticus, e.g., complex partial non-convulsivestatus epilepticus, simple partial non-convulsive status epilepticus,subtle non-convulsive status epilepticus; generalized non-convulsivestatus epilepticus, e.g., late onset absence non-convulsive statusepilepticus, atypical absence non-convulsive status epilepticus, ortypical absence non-convulsive status epilepticus.

Seizure

A seizure is the physical findings or changes in behavior that occurafter an episode of abnormal electrical activity in the brain. The term“seizure” is often used interchangeably with “convulsion.” Convulsionsare when a person's body shakes rapidly and uncontrollably. Duringconvulsions, the person's muscles contract and relax repeatedly.

Based on the type of behavior and brain activity, seizures are dividedinto two broad categories: generalized and partial (also called local orfocal). Classifying the type of seizure helps doctors diagnose whetheror not a patient has epilepsy.

Generalized seizures are produced by electrical impulses from throughoutthe entire brain, whereas partial seizures are produced (at leastinitially) by electrical impulses in a relatively small part of thebrain. The part of the brain generating the seizures is sometimes calledthe focus.

There are six types of generalized seizures. The most common anddramatic, and therefore the most well-known, is the generalizedconvulsion, also called the grand-mal seizure. In this type of seizure,the patient loses consciousness and usually collapses. The loss ofconsciousness is followed by generalized body stiffening (called the“tonic” phase of the seizure) for 30 to 60 seconds, then by violentjerking (the “clonic” phase) for 30 to 60 seconds, after which thepatient goes into a deep sleep (the “postictal” or after-seizure phase).During grand-mal seizures, injuries and accidents may occur, such astongue biting and urinary incontinence.

Absence seizures cause a short loss of consciousness (just a fewseconds) with few or no symptoms. The patient, most often a child,typically interrupts an activity and stares blankly. These seizuresbegin and end abruptly and may occur several times a day. Patients areusually not aware that they are having a seizure, except that they maybe aware of “losing time.”

Myoclonic seizures consist of sporadic jerks, usually on both sides ofthe body. Patients sometimes describe the jerks as brief electricalshocks. When violent, these seizures may result in dropping orinvoluntarily throwing objects.

Clonic seizures are repetitive, rhythmic jerks that involve both sidesof the body at the same time.

Tonic seizures are characterized by stiffening of the muscles.

Atonic seizures consist of a sudden and general loss of muscle tone,particularly in the arms and legs, which often results in a fall.

Seizures described herein can include epileptic seizures; acuterepetitive seizures; cluster seizures; continuous seizures; unremittingseizures; prolonged seizures; recurrent seizures; status epilepticusseizures, e.g., refractory convulsive status epilepticus, non-convulsivestatus epilepticus seizures; refractory seizures; myoclonic seizures;tonic seizures; tonic-clonic seizures; simple partial seizures; complexpartial seizures; secondarily generalized seizures; atypical absenceseizures; absence seizures; atonic seizures; benign Rolandic seizures;febrile seizures; emotional seizures; focal seizures; gelastic seizures;generalized onset seizures; infantile spasms; Jacksonian seizures;massive bilateral myoclonus seizures; multifocal seizures; neonatalonset seizures; nocturnal seizures; occipital lobe seizures; posttraumatic seizures; subtle seizures; Sylvan seizures; visual reflexseizures; or withdrawal seizures. In some embodiments, the seizure is ageneralized seizure associated with Dravet Syndrome, Lennox-GastautSyndrome, Tuberous Sclerosis Complex, Rett Syndrome or PCDH19 FemalePediatric Epilepsy.

EXAMPLES

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. The synthetic andbiological examples described in this application are offered toillustrate the compounds, pharmaceutical compositions, and methodsprovided herein and are not to be construed in any way as limiting theirscope. In the synthetic examples below, the descriptions of experimentalprocedures within a reaction sequence are listed in numerical order.

In some cases, the stereochemistry assigned herein (e.g., the assignmentof “R” or “S” to the C22 position of the steroid) may be tentatively(e.g., randomly) assigned. For example, a C22 position may be drawn inthe “R” configuration when the absolute configuration is “S.” A C22position may also be drawn in the “S” configuration when the absoluteconfiguration is “R.” Such random assignment applies to compounds 7, 13,14, 18, 19, 22, 25, 27, 31, 37, 41, 50, 55, 60, 63, 66, 68, 73, 79, 86,89, 91, and 99.

Materials and Methods

The compounds provided herein can be prepared from readily availablestarting materials using the following general methods and procedures.It will be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and P. G. M. Wuts, ProtectingGroups in Organic Synthesis, Second Edition, Wiley, New York, 1991, andreferences cited therein.

The compounds provided herein may be isolated and purified by knownstandard procedures. Such procedures include (but are not limited to)recrystallization, column chromatography, HPLC, or supercritical fluidchromatography (SFC). The following schemes are presented with detailsas to the preparation of representative pyrazoles that have been listedherein. The compounds provided herein may be prepared from known orcommercially available starting materials and reagents by one skilled inthe art of organic synthesis. Exemplary chiral columns available for usein the separation/purification of the enantiomers/diastereomers providedherein include, but are not limited to, CHIRALPAK® AD-10, CHIRALCEL® OB,CHIRALCEL® OB-H, CHIRALCEL® OD, CHIRALCEL® OD-H, CHIRALCEL® OF,CHIRALCEL® OG, CHIRALCEL® OJ and CHIRALCEL® OK.

Exemplary general method for preparative HPLC: Column: Waters RBridgeprep 10 μm C18, 19*250 mm. Mobile phase: acetonitrile, water (NH₄HCO₃)(30 L water, 24 g NH₄HCO₃, 30 mL NH₃.H₂O). Flow rate: 25 mL/min

Exemplary general method for analytical HPLC: Mobile phase: A: water (10mM NH₄HCO₃), B: acetonitrile Gradient: 5%-95% B in 1.6 or 2 min Flowrate: 1.8 or 2 mL/min; Column: XBridge C18, 4.6*50 mm, 3.5 μm at 45° C.

NMDA Modulation

NMDA potentiation in mammalian cells which expressed NMDA receptors wasassessed using the automatic patch-clamp system can be used determinethe NAM activity of compounds as described below. An whole cellpatch-clamp system can be used determine the PAM activity of compoundsas described below.

Automated Patch-Clamp System (QPatch HTX):

In this study, HEK 293 cells stably transfected with glutamate-activatedchannels of the GRIN1/2A subtype will be used together with submaximalNMDA concentrations (300 μM NMDA, co-application with 8 μM Glycine) toinvestigate the negative allosteric modulation of the test compounds.Percent potentiation results obtained with this method are shown inTable 1.

Cell Culture

In general, cells will be passaged at a confluence of about 80% to 90%.For electrophysiological measurements cells will be harvested at aconfluence of about 80% to 90% from sterile culture flasks containingculture complete medium. Cells will be transferred as suspension in PBSto the QPatch 16X or QPatch HTX system to the centrifuge/washerdirectly.

Standard Laboratory Conditions: Cells will be incubated at 37° C. in ahumidified atmosphere with 5% CO₂ (rel. humidity about 95%).

Culture media: The cells will be continuously maintained in and passagedin sterile culture flasks containing a 1:1 mixture of Dulbecco'smodified eagle medium and nutrient mixture F-12 (D-MEM/F-12 1×, liquid,with L-Glutamine) supplemented with 10% fetal bovine serum, 1%Penicillin/Streptomycin solution, and 50 μM AP-5 blocker.

Antibiotics: The complete medium as indicated above is supplemented with100 μg/mL hygromycin, 15 μg/mL blasticidin and 1 μg/mL puromycin.

Induction of Expression: 2.5 μg/mL tetracycline is added 24 h beforestart of experiments.

Dose Formulation

Dose levels are in terms of test compounds, as supplied. Vehicle will beadded to achieve a stock concentration of 10 mM (storage at −10° C. to−30° C.). A further stock solutions of 1.0 mM will be prepared in DMSO.Details of stock solution usage (thawing, dose formulations) will bedocumented in the raw data. The time period of stock solution usage willbe detailed in the report.

Test Compound Concentrations

Dose levels are in terms of test compounds, as supplied. Vehicle will beadded to achieve a stock concentration of 10 mM (storage at −10° C. to−30° C.). A further stock solutions of 1.0 mM will be prepared in DMSO.Details of stock solution usage (thawing, dose formulations) will bedocumented in the raw data. The time period of stock solution usage willbe detailed in the report.

One test concentration of 1.0 μM will be tested.

All test solutions will be prepared by diluting the stock solutions witheither Mg-free bath solution only or Mg-free bath solution containingNMDA (300 μM) and glycine (8.0 μM) shortly prior to theelectrophysiological experiments and kept at room temperature (19° C. to30° C.) when in use. 0.1% DMSO will be used as vehicle.

Frequency of preparation: For each test concentration, fresh solutionsof test compounds will be prepared every day.

Stability of dose formulation: All preparation times will be documentedin the raw data. Any observations regarding instability of testcompounds will be mentioned in the raw data.

Storage of dose formulation: On the day of experimentation doseformulations will be maintained at room temperature (19° C. to 30° C.)when in use.

Bath Solutions

For preparing the experiments and for formation of the giga-ohm-seal,the following standard bath solution will be used:

Sodium Chloride: 137 mM; Potassium Chloride: 4 mM; Calcium Chloride: 1.8mM; Magnesium Chloride: 1 mM; HEPES: 10 mM; D-Glucose: 10 mM; Cremophor:0.02%; pH (NaOH): 7.4

The 1× bath solution will be prepared by diluting 10× bath solutionwithout Glucose and 100× Glucose solution with water at least every 7days. Both stock solutions have been prepared prior to the experimentalstart of the present study and stored at 1° C. to 9° C. (10× bathsolution) or −10° C. to −30° (100× Glucose solution). The batchnumber(s) of the bath solution(s) used in the experiments will bedocumented in the raw data. When in use, the 1× bath solution will bekept at room temperature (19° C. to 30° C.). When not in use, the 1×bath solution will be stored at 1° C. to 9° C.

After the giga-seal was formed the following Mg-free bath solution willbe used: Sodium Chloride: 137 mM; Potassium Chloride: 4 mM; CalciumChloride; 2.8 mM; HEPES: 10 mM; D-Glucose: 10 mM; Cremophor: 0.02%; pH(NaOH): 7.4 This Mg-free bath solution will be prepared as a 1× solutionand stored at 1° C. to 9° C. It will be prepared freshly at least every10 days.

Intracellular Solution

The 1× intracellular solution will be thawed every day out of a frozen1× intracellular solution, which has been prepared prior to theexperimental start of the present study, aliquoted and stored at −10° C.to −30° C. When in use, the 1× intracellular solution will kept at roomtemperature (19° C. to 30° C.). Remaining 1× intracellular solution willbe stored in the fridge (1° C. to 9° C.). The 1×intracellular solutionwill include the components outlined below:

Potassium Chloride: 130 mM; Magnesium Chloride: 1 mM; Mg-ATP: 5 mM;HEPES: 10 mM; EGTA: 5 mM; pH (KOH): 7.2

Cell Treatment

For this study, cells will continuously be perfused with NMDA/Glycine,Test Compound or Test Compound/NMDA/Glycin.

In every case, at least 30-second prewash steps with a test compoundwill be performed in between applications. For details see Table Abelow.

Each experiment type will be analyzed in at least n=3 isolated cells.The NMDA and Glycine stock solutions will be prepared prior to theexperimental start of the present study, stored frozen (−10° C. to −30°C.) until the day of experimentation. Shortly prior to theelectrophysiological experiments, frozen stock solutions will be thawedand diluted.

Control: The effect of vehicle (0.1% DMSO) andD-(−)-2-Amino-5-phosphonopentanoic acid (AP-5) (100 μM) will be measuredat three cells every second week, in order to assure successfulexpression of NMDA receptors.

The 50 mM stock solution of AP-5 has been prepared prior to theexperimental start of the present study, aliquoted and stored frozen(−10° C. to −30° C.) until the day of experimentation. Shortly prior tothe electrophysiological experiments the frozen stock solution will bethawed and then diluted in Mg-free bath solution containing NMDA (300μM) and glycine (8.0 μM), to give a final perfusion concentration of 100μM.

Experimental Procedure

Cells are transferred as suspension in serum-free medium to the QPatchHTX system and kept in the cell storage tank/stirrer during experiments.All solutions applied to cells including the intracellular solution willbe maintained at room temperature (19° C. to 30° C.).

During the sealing process standard bath solution described above willbe used. All solutions applied to cells including the pipette solutionwill be maintained at room temperature (19° C. to 30° C.). Afterformation of a Gigaohm seal between the patch electrodes and transfectedindividual HEK293 cells only Mg-free bath solution will be perfused andthe cell membrane will be ruptured to assure electrical access to thecell interior (whole-cell patch-configuration). Inward currents will bemeasured upon application of 300 μM NMDA (and 8.0 μM Glycine) topatch-clamped cells for 5 sec. During the entire experiment the cellswill be voltage-clamped at a holding potential of −80 mV.

For the analysis of test compounds, NMDA receptors will be stimulated by300 μM NMDA and 8.0 μM Glycine and test compound combinations describedbelow. Thirty-second prewash steps with a test compound will beperformed in between applications.

TABLE A Application Protocol; use dependence of test compounds Appl. #Duration(s) Application 1 4 NMDA/Glycine 2 30 Bath 3 4 NMDA/Glycine 2repetitions 4 30 1 μM Test Compound 5 4 1 μM Test Compound +NMDA/Glycine 6 repetitions 6 30 Bath 7 4 NMDA/Glycine 2 repetitions

TABLE B Application Protocol; control experiments Appl. # Duration(s)Application 1 4 NMDA/Glycine 2 30 Bath 3 4 NMDA/Glycine 2 repetitions 430 Bath 5 4 NMDA/Glycine 6 repetitions 6 30 Bath 7 4 NMDA/Glycine + 100μM AP-5 2 repetitionsWhole-Cell Patch Clamp of Mammalian Cells (Ionworks Barracuda (IWB)):

The whole-cell patch-clamp technique was used to investigate the effectsof positive allosteric modulating activity of test compoundson_GlunN1/GluN2A and GluN2B glutamate receptors expressed in mammaliancells. EC₅₀ and E_(max) data are shown in Table 1.

HEK293 cells were transformed with adenovirus 5 DNA and transfected withcDNA encoding the human GRIN1/GRIN2A genes. Stable transfectants wereselected using G418 and Zeocin-resistance genes incorporated into theexpression plasmid and selection pressure maintained with G418 andZeocin in the medium. Cells were cultured in Dulbecco's Modified EagleMedium/Nutrient Mixture (D-MEM/F-12) supplemented with 10% fetal bovineserum, 100 μg/ml penicillin G sodium, 100 μg/ml streptomycin sulphate,100 μg/ml Zeocin, 5 μg/ml blasticidin and 500 μg/ml G418.

Test article effects were evaluated in 8-point concentration-responseformat (4 replicate wells/concentration). All test and control solutionscontained 0.3% DMSO and 0.01% Kolliphor® EL (C5135, Sigma). The testarticle formulations were loaded in a 384-well compound plate using anautomated liquid handling system (SciClone ALH3000, CaliperLifeScienses). The measurements were performed using Ion Works Barracudaplatform following this procedure:

Electrophysiological Procedures:

-   -   a) Intracellular solution (mM): 50 mM CsCl, 90 mM CsF, 2 mM        MgCl₂, 5 mM EGTA, 10 mM HEPES. Adjust to pH 7.2 with CsOH.    -   b) Extracellular solution, HB-PS (composition in mM): NaCl, 137;        KCl, 1.0; CaCl₂, 5; HEPES, 10; Glucose, 10; pH adjusted to 7.4        with NaOH (refrigerated until use).    -   c) Holding potential: −70 mV, potential during agonist/PAM        application: −40 mV.        Recording Procedure:    -   a) Extracellular buffer will be loaded into the PPC plate wells        (11 μL per well). Cell suspension will be pipetted into the        wells (9 μL per well) of the PPC planar electrode.    -   b) Whole-cell recording configuration will be established via        patch perforation with membrane currents recorded by on-board        patch clamp amplifiers.    -   c) Two recordings (scans) will be performed. First, during        pre-application of test article alone (duration of        pre-application—5 min) and second, during test articles and        agonist (EC₂₀ L-glutamate and 30 μM glycine) co-application to        detect positive modulatory effects of the test article.

Test Article Administration: The first pre-application will consist ofthe addition of 20 μL of 2× concentrated test article solution and,second, of 20 μL of 1× concentrated test article and agonist at 10 μL/s(2 second total application time).

Potentiating Effect of Positive Allosteric Modulators (PAM) on theChannel

Potentiating effect of positive allosteric modulators (PAM) on thechannel will be calculated as% activation=(I _(PAM) /I _(EC10-30))×100%−100%where I_(PAM) will be the L-glutamate EC₁₀₋₃₀—elicited current inpresence of various concentrations of test articles and I_(EC20) will bethe mean current elicited with L-glutamate EC₂₀. PAMconcentration-response data will be fitted to an equation of the form:% Activation=% L-glutamate EC₂₀+{(% MAX−% L-glutamateEC₂₀)/[1+([Test]/EC₅₀)^(N)]},where [Test] will be the concentration of PAM (test article), EC₅₀ willbe the concentration of PAM producing half-maximal activation, N will bethe Hill coefficient, % L-glutamate EC₂₀ will be the percentage of thecurrent Elicited with L-glutamate EC₂₀, % MAX is the percentage of thecurrent activated with the highest dose of PAM co-admitted withL-glutamate EC₂₀ and % Activation will be the percentage of the currentelicited with L-glutamate EC₁₀₋₃₀ at each PAM concentration.

The maximal amplitude of the evoked currents are measured and defined asPeak Current Amplitude (PCA).

Abbreviations

PCC: pyridinium chlorochromate; t-BuOK: potassium tert-butoxide; 9-BBN:9-borabicyclo[3.3.1]nonane; Pd(t-Bu₃P)₂:bis(tri-tert-butylphosphine)palladium(0); AcCl: acetyl chloride;i-PrMgCl: Isopropylmagnesium chloride; TBSCl:tert-Butyl(chloro)dimethylsilane; (i-PrO)₄Ti: titaniumtetraisopropoxide; BHT: 2,6-di-t-butyl-4-methylphenoxide; Me: methyl;i-Pr: iso-propyl; t-Bu: tert-butyl; Ph: phenyl; Et: ethyl; Bz: benzoyl;BzCl: benzoyl chloride; CsF: cesium fluoride; DCC:dicyclohexylcarbodiimide; DCM: dichloromethane; DMAP:4-dimethylaminopyridine; DMP: Dess-Martin periodinane; EtMgBr:ethylmagnesium bromide; EtOAc: ethyl acetate; TEA: triethylamine; AlaOH:alanine; Boc: t-butoxycarbonyl. Py: pyridine; TBAF:tetra-n-butylammonium fluoride; THF: tetrahydrofuran; TBS:t-butyldimethylsilyl; TMS: trimethylsilyl; TMSCF₃:(Trifluoromethyl)trimethylsilane; Ts: p-toluenesulfonyl; Bu: butyl;Ti(OiPr)₄: tetraisopropoxytitanium; LAH: Lithium Aluminium Hydride; LDA:lithium diisopropylamide; LiOH.H₂O: lithium hydroxide hydrates; MAD:methyl aluminum bis(2,6-di-t-butyl-4-methylphenoxide); MeCN:acetonitrile; NBS: N-bromosuccinimide; Na₂SO₄: sodium sulfate; Na₂S₂O₃:sodium thiosulfate; PE: petroleum ether; MeCN: acetonitrile; MeOH:methanol; Boc: t-butoxycarbonyl; MTBE: methyl tert-butyl ether; DIAD:diisopropyl azodicarboxylate; sat.: saturated; aq.: aqueous; hr/hrs:hour/hours; min/mins: minute/minutes.

Example 1: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-3-(trifluoromethyl)-2,3,4,7,8,9,10,11,12,13,14,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(1)

1. To a solution of TBAF (3.04 mL, 1 M in THF, 3.04 mmol, Aldrich) inTHF (100 mL) was added TMSCF₃ (25.8 g, 182 mmol) followed by a solutionof S-200-INT-2 (19 g, 60.8 mmol) in THF (100 mL) dropwise at 0° C. Themixture was stirred at 0° C. for 30 min. To the mixture was added TBAF(200 mL, 1 M in THF, 200 mmol, domestic) at 0° C. The mixture wasstirred at 0° C. for another 30 mins. To the mixture was added NH₄Cl(100 mL, sat., aq.). The mixture was concentrated under vacuum. To theresidue was added PE/EtOAc (400 mL, 1:1), the organic layer wasseparated, which was combined with other two batches (2×10 g ofS200-INT-2). The combined organic layer was washed with water (300 mL),brine (300 mL), dried over Na₂SO₄, filtered and concentrated in vacuumto give an oil. The residue was dissolved in DCM (150 mL) and dilutedwith PE (750 mL). The solution was poured into a silica gel column (500g, 100-200 mesh) and eluted with PE:DCM:EtOAc=5:1:0.05 to 5:1:0.1 togive S200-CF3_1B (12 g, 70% purity, 17% yield) as an oil and impureS200-CF3_1A. The impure was re-crystallized from MeCN (250 mL) to giveS200-CF3_1A (6.5 g) as a solid. The filtrated form MeCN was purified bysilica gel column (PE:DCM:EtOAc=50:1:1 to 20:1:1) to give a crude whichwas re-crystallized from MeCN (20 mL) to give S-200-CF3_1A (1 g, 16%total yield) as a solid.

Note: 200-CF3_1A and 200-CF3_1B were identified from ³J_(H,CF) ₃ (FDCS).(J. Org. Chem. 2015, 80, 1754.

S-200-CF3_1A:

¹H NMR (400 MHz, CDCl₃) δ 5.43-5.33 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H);2.49 (s, 2H); 2.11-1.97 (m, 4H), 1.95-1.32 (m, 14H), 1.30-0.98 (m, 7H),0.59 (s, 3H).

S-200-CF3_1B:

¹H NMR (400 MHz, CDCl₃) δ 5.54-5.41 (m, 1H), 4.86 (s, 1H), 4.72 (s, 1H);2.78-2.65 (m, 1H); 2.18-1.97 (m, 3H), 1.95-1.35 (m, 16H), 1.32-0.98 (m,7H), 0.59 (s, 3H). 2. To a solution of S-200-CF3_1A (8 g, 20.9 mmol) inTHF (80 mL) was added 9-BBN dimer (5.85 g, 24 mmol). The mixture wasstirred at 40° C. for 1 h. The mixture was cooled to 0° C. To themixture was added EtOH (12 mL), NaOH (41.8 mL, 5 M, aq.) and H₂O₂ (20.9mL, 10 M, aq.) dropwise. The mixture was stirred at 50° C. for 1 h. Tothe mixture was added Na₂SO₃ (100 mL, 25%, aq.) after cooling. Themixture was extracted with EtOAc (300 mL). The organic layer wasseparated and purified by silica gel column (PE:EtOAc=10:1 to 5:1) togive S-200-CF3_2A (7.1 g, 85%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.42-5.32 (m, 1H), 3.64 (dd, J=3.2, 10.4 Hz,1H), 3.37 (dd, J=6.8, 10.4 Hz, 1H), 2.49 (s, 2H), 2.32-1.92 (m, 4H),1.92-1.70 (m, 4H), 1.70-1.29 (m, 8H), 1.29-0.91 (m, 11H), 0.71 (s, 3H).

3. DMP (6.31 g, 14.9 mmol) was added to a solution of S-200-CF3_5A (3 g,7.49 mmol) in DCM (50 mL) at 25° C., after stirring at 25° C. for 30min, the reaction mixture was quenched with saturated NaHCO₃ (100 mL)and DCM (100 mL) was added and stirred for 10 min. The DCM phase wasseparated and washed with saturated aqueous Na₂S₂O₃ (2×100 mL). Thecombined organic layer was washed with saturated brine (2×100 mL), driedover anhydrous Na₂SO₄, filtered and concentrated. The residue waspurified by flash column (5˜20% of EtOAc in PE) to give N-004-027_1 (1.5g, 50%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.58-9.55 (m, 1H), 5.38-5.36 (m, 1H), 2.49 (s,1H), 2.40-2.25 (m, 1H), 2.23-1.60 (m, 10H), 1.53-1.20 (m, 9H), 1.15-1.00(m, 7H), 0.78-0.64 (m, 3H).

4. To a solution of N-004-027_1 (1.5 g, 3.76 mmol) in anhydrous THF (40mL) was added CsF (1.42 g, 9.40 mmol) at 0° C. After stirring at 0° C.for 20 min, TMSCF₃ (1.33 g, 9.40 mmol) was added at 0° C. and stirredfor 30 min. The color becomes light yellow. TBAF.3H₂O (4.74 g, 15.0mmol) was added and stirred at 50° C. for 30 min. The reaction mixturewas poured into ice-water (100 mL). The aqueous phase was extracted withEtOAc (2×100 mL). The combined organic phase was washed with saturatedbrine (2×100 mL), dried over anhydrous Na₂SO₄, filtered and concentratedto give a mixture of isomers (1.45 g, crude) as a yellow solid, whichwas purified by flash column (0˜15% of EtOAc in PE) to give 53 (340 mg,24%) as a white solid and 1 (200 mg, 14%) as a white solid. 1:

¹H NMR (400 MHz, CDCl₃) δ 5.38-5.36 (m, 1H), 4.10-4.00 (m, 1H), 2.49 (s,2H), 2.19-2.12 (m, 1H), 2.06-1.61 (m, 10H), 1.53-1.29 (m, 6H), 1.27-0.98(m, 10H), 0.71 (s, 3H).

LCMS Rt=1.121 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,

MS 50-100_1_4 min·m, for C₂₄H₃₃F₆O [M+H-H₂O]⁺ 451, found 451.

1:

¹H NMR (400 MHz, CDCl₃) δ 5.38-5.36 (m, 1H), 4.10-4.00 (m, 1H), 2.49 (s,2H), 2.19-2.12 (m, 1H), 2.06-1.61 (m, 10H), 1.53-1.29 (m, 6H), 1.27-0.98(m, 10H), 0.71 (s, 3H).

LCMS Rt=1.121 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,

MS 50-100_1_4 min·m, for C₂₄H₃₃F₆O [M+H-H₂O]⁺ 451, found 451.

Example 2: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3-(methoxymethyl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(2)

1. To a suspension of N-4-4_1 (50 g, 157 mmol) in anhydrous methanol(500 mL) at 20° C. was added anhydrous TsOH (2.84 g, 15.7 mmol) in oneportion. The mixture was warmed up to 60° C. and stirred for 1 h. Thereaction mixture was quenched with Et₃N (1.58 g, 15.7 mmol) and stirredfor an additional 30 min. The precipitated solid was filtered out,washed with methanol (250 mL) and dried in air to give N-4-1_2 (51 g,90%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.18 (s, 3H), 3.14 (s, 3H); 2.54-2.48 (m, 1H);2.10-2.00 (m, 4H); 1.95-1.75 (m, 2H), 1.65-1.50 (m, 7H), 1.48-0.80 (m,11H), 0.78-0.75 (m, 4H), 0.59 (s, 3H).

2. To a suspension of Ph₃PMeBr (75 g, 210 mmol) in anhydrous THF (500mL) under N₂ at 20° C. was added t-BuOK (23.5 g, 210 mmol) in portions.The mixture became deep orange and stirred at 20° C. for 30 mins. ThenN-4-1_2 (51 g, 140 mmol) was added. The mixture was warmed to 40° C. andstirred for 1 h. The reaction mixture was cooled and poured into aqueousNH₄Cl (ice)(400 mL) in portions. The resulting mixture was separated;the aqueous layer was extracted with THF (200 mL). The combined organiclayer was used as a solution of N-4-1_3 directly without furtherpurification.

3. To a solution of N-4-1_3 (50.4 g, 139 mmol) in THF (700 mL) was addedaqueous HCl (1 M, 208 mL, 208 mmol) at 20° C. The mixture was stirred at20° C. for 1 hr, and a solid precipitated. Water (200 mL) was added tothe mixture, and the precipitated solid was filtered out, washed withwater and dried to give N-4-1_4 (41 g, 94%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.85 (s, 1H), 4.70 (s, 1H); 2.38-2.25 (m, 3H);2.10-1.98 (m, 3H), 1.88-1.49 (m, 10H), 1.40-1.08 (m, 11H), 0.97-0.72 (m,2H), 0.58 (s, 3H).

4. To a solution of Me₃SI (101 g, 496 mmol) in anhydrous THF (400 mL) at25° C. under N₂ was added t-BuOK (58.3 g, 520 mmol) in portions andstirred for 30 mins. A solution of N-4-1_4 (39 g, 124 mmol) in anhydrousTHF (300 mL) was added. The reaction mixture was warmed to 50° C. andstirred for 2 hrs. The reaction mixture was cooled to 25° C. and wastreated with aq. NH₄Cl (500 mL). The aqueous phase was extracted withEtOAc (2×500 mL). The combined organic phase was washed with brine(2×300 mL), dried over anhydrous Na₂SO₄, filtered and concentrated undervacuum. The residue was purified by silica gel chromatography(PE/EtOAc=20/1 to 10/1) to afford N-4-3_1 (35 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H); 2.65-2.55 (m, 2H);2.10-1.98 (m, 2H), 1.92-1.49 (m, 13H), 1.40-1.13 (m, 8H), 0.99-0.69 (m,6H), 0.57 (s, 3H).

5. To a solution of N-4-3_1 (35 g, 647 mmol) in anhydrous MeOH (500 mL)was added MeONa (57.2 g, 1.06 mol) at 25° C. and the mixture was stirred30 min under N₂. The reaction mixture was warmed to 70° C. and stirredat reflux for 3 hrs under N₂. The reaction mixture was cooled to 25° C.and treated with water (500 mL). The aqueous phase was extracted withDCM (2×300 mL). The combined organic phase was washed with saturatedbrine (2×300 mL), dried over anhydrous Na₂SO₄, filtered and concentratedin vacuum to obtain a solid. The residue was purified by silica gelchromatography (PE/EtOAc=10/1 to 6/1) to afford N-4-3_2 (25 g, impure)as a solid. The crude product was triturated from PE (250 mL) at 25° C.for 1 h. The suspension was filtered and the filter cake was dried undervacuum to obtain N-4-3_2 (15 g, 25%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.86 (s, 1H), 4.72 (s, 1H), 3.46-3.37 (m, 5H),2.54 (s, 1H), 2.07-1.99 (m, 1H), 1.89-1.52 (m, 15H), 1.41-1.06 (m, 10H),0.86 (s, 3H); 0.58 (s, 3H)

6. To a solution of N-4-3_2 (15 g, 41.6 mmol) in anhydrous THF (200 mL)was added 9-BBN dimer (27.7 g, 124 mmol) at 0° C. and stirred for 30mins under N₂. The reaction mixture was warmed to 50° C. and stirred for1 h. The reaction mixture was cooled to 0° C. and EtOH (50 mL) wasadded, then NaOH (41.6 mL, 5 M, 208 mmol) at 0° C. was added veryslowly. H₂O₂ (23.5 g, 208 mmol, 30% in water) was added slowly whilekeeping the inner temperature below 10° C. The mixture was warmed to 50°C. and stirred for another 1 h. The reaction mixture was cooled, pouredinto ice-water (500 mL) in portions and filtered. The filtrate wasconcentrated under vacuum to provide N-4-3_3 (14 g, crude) as an oil.The crude residue was used directly for the next step. 7. DMP (3.35 g,7.92 mmol) was added to a mixture of N-4_3 (1 g, 2.64 mmol) in DCM (20mL) at 25° C. The reaction mixture was warmed to 40° C. and stirred for1 h. The reaction mixture was quenched with saturated aqueous NaHCO₃ atpH 7-8 and below 10° C. The suspension was filtered. The DCM phase inthe filtrate was separated and washed with saturated NaHCO₃/Na₂S₂O₃aqueous (1:1, 2×50 mL), brine (2×50 mL), dried over Na₂SO₄, filtrate andconcentrated in vacuum to obtain a solid. The residue was purified byflash column (0-30% of EtOAc in PE) to give N-4-3_4 (0.6 g, 60%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 9.57 (s, 1H), 3.40-3.34 (m, 5H); 2.38-2.28 (m,1H); 1.94-1.76 (m, 2H), 1.74-1.35 (m, 16H), 1.06-0.82 (m, 10H),0.73-0.64 (m, 5H).

8. Isopentylmagnesium bromide (4.37 mL, 8.74 mmol 2 M in diethyl ether)was added to a solution of N-4-3_4 (0.6 g, 1.59 mmol) in anhydrous THF(10 mL) at 0° C. under N₂. The reaction mixture was warmed to 25° C. andstirred for 1 hour. To the reaction mixture was added saturated aqueousNH₄Cl (50 mL) solution. The aqueous phase was extracted with EtOAc (3×50mL). The combined organic phase was washed with saturated brine (2×50mL), dried over anhydrous Na₂SO₄, filtered and concentrated in to getN-4-4A (0.5 crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.64-3.60 (m, 1H), 3.40-3.37 (m, 5H);2.02-1.79 (m, 3H); 1.75-1.50 (m, 11H), 1.25-1.10 (m, 14H), 0.99-0.75 (m,14H), 0.70-0.64 (m, 4H).

9. DMP (1.88 g, 4.44 mmol) was added to a solution of N-4-4A (0.5 g,crude) in DCM (20 mL) at 25° C. The reaction mixture was warmed to 40°C. and stirred for 1 hr. The reaction mixture was quenched withsaturated NaHCO₃ aqueous at pH 7˜8 and below 10° C. The suspension wasfiltered. The DCM phase was separated and washed with saturatedNaHCO₃/Na₂S₂O₃ aqueous (1:1, 2×50 mL), brine (2×50 mL), dried overNa₂SO₄ filtered and concentrated under vacuum to get N-4-4O (0.4 g,crude) as a solid, which was used directly for the next step. NaBH₄(0.340 g, 8.95 mmol) was slowly added to a solution of N-4-4O (0.4 g,0.895 mmol) in MeOH (4 mL) was slowed added at 25° C. and stirred for 2hrs. The aqueous phase was extracted with DCM (2×20 mL). The combinedorganic phase was washed with saturated brine (2×20 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under vacuum to obtain asolid. The residue was purified by silica gel chromatography(PE/EtOAc=8/1 to 5/1) to afford 35 (150 mg, impure) and 2 (130 mg,impure) as solids. 2 (130 mg, impure) was re-crystallized from MeCN (3mL) at 82° C. reflux for 1 hr. The mixture was stirred and cooled to 25°C. The suspension was filtered and the filtrate concentrated undervacuum to provide 2 (50 mg, 12%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.63-3.61 (m, 1H), 3.41-3.38 (m, 5H); 2.51 (s,1H); 1.97-1.81 (m, 2H), 1.71-1.31 (m, 15H), 1.26-1.03 (m, 10H),0.97-0.78 (m, 14H), 0.71-0.59 (m, 4H).

LCMS Rt=1.350 min in 2.0 min chromatography, 30-90 AB, purity 99%, MSESI calcd. for C₂₉H₄₈O [M+H-2H₂O]⁺ 413, found 413.

Example 3: Synthesis of(3S,8R,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-13-methyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(3)

1. t-BuOH (350 mL) was charged into a three-neck round bottom flaskunder nitrogen at 35° C. and stirred under nitrogen gas for 10 mins.t-BuOK (90.5 g, 807 mmol) was added to the mixture and stirred undernitrogen gas for 15 mins. S-310-B9_1 (20 g, 73.4 mmol) was added to theabove mixture and stirred under nitrogen gas at 35° C. for 1.5 hrs. Thereaction mixture was poured into 10% aqueous acetic acid (500 mL) andstirred for 15 mins and below 35° C. Water (500 mL) was added and themixture was stirred for 30 mins. The pH of the mixture was adjusted to7-8 with sodium bicarbonate (500 ml) and stirred for 30 mins. Themixture was extracted with PE (2×500 mL). The organic layer wasseparated, washed with brine (500 mL), dried over anhydrous sodiumsulfate, filtered and concentrated below 35° C. to give S-200-N19-3_1(17 g, crude) as an oil. The crude residue was used directly for thenext step.

2. To a solution of 2,6-di-tert-butyl-4-methylphenol (100 g, 453 mmol)in toluene (300 ml) was added, drop-wise, AlMe₃ (113 mL, 226 mmol, 2 Min toluene) at 0° C. The mixture was stirred at 25° C. for 1 hr togenerate MAD. A solution of S-200-N19-3_1 (10 g, 36.7 mmol) in toluene(50 mL) was added dropwise to the MAD solution at −70° C. After stirringat −70° C. for 1 hour, MeMgBr (36.6 ml, 110 mmol, 3M in ethyl ether) wasadded drop wise at −70° C. The resulting solution was stirred at −70° C.for 1 hr. The reaction mixture was quenched by saturated citric acid(400 ml) at −70° C. After stirring at 25° C. for 10 min, the resultingmixture was filtered and washed with EtOAc (2×200 ml). The combinedorganic layer was separated, washed with brine (2×200 ml), dried overNa₂SO₄, filtered and concentrated under vacuum. The residue was purifiedby silica gel chromatography (PE/EtOAc=10/1 to 5/1) to yieldS-200-N19-3_2 (7.6 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.45-5.40 (m, 1H), 2.51-2.38 (m, 1H),2.49-2.21 (m, 1H), 2.14-1.88 (m, 5H), 1.86-1.77 (m, 2H), 1.73-1.38 (m,8H), 1.34-1.22 (m, 4H), 0.95-0.81 (m, 8H).

3. To a suspension of PPh₃EtBr (37.1 g, 100 mmol) in THF (200 mL) underN₂ was added t-BuOK (11.2 g, 100 mmol) at 40° C. After stirring at 20°C. for 10 min, S-200-N19-3_2 (7.6 g, 25.1 mmol) was added. The reactionmixture was stirred at 40° C. for 1 hour. The reaction was quenched withaq.NH₄Cl (200 mL) at 0° C., extracted with EtOAc (3×200 mL). Thecombined organic phase was washed with brine (200 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified byCombi-flash (0%˜30% of EtOAc in PE) to afford S-200-N19-3_3 (5 g, 63%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.45-5.35 (m, 1H), 5.20-5.00 (m, 1H),2.41-2.30 (m, 1H), 2.29-2.12 (m, 3H), 2.09-1.76 (m, 6H), 1.69-1.38 (m,15H), 1.35-0.94 (m, 7H).

4. To a solution of S-200-N19-3_3 (2 g, 6.35 mmol) in THF (20 mL) wasadded 9-BNN dimer (3.09 g, 12.7 mmol) at 0° C. under N₂. The solutionwas stirred at 60° C. for 1 hr. After cooling to 0° C., a solution ofEtOH (20 ml) and NaOH (12.7 ml, 5M, 63.5 mmol) was added very slowly.After addition, H₂O₂ (2.15 mg, 6.35 mmol, 30% in water) was added slowlyand the inner temperature was maintained below 10° C. The mixture wasstirred at 60° C. under N₂ for 1 hr. The mixture was re-cooled to 30° C.Water (100 mL) was added to the solution and the aqueous layer extractedwith EtOAc (100 mL). The organic layer was washed with brine (2×100 mL).The combined organic layer was dried over anhydrous Na₂SO₄, and waspurified by silica gel chromatography (PE/EtOAc=2/1) to affordS-200-N19-4_1 (1.6 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.45-5.35 (m, 1H), 3.75-3.62 (m, 1H),2.28-2.19 (m, 1H), 2.10-1.75 (m, 7H), 1.71-0.97 (m, 19H), 0.92-0.75 (m,4H), 0.68 (s, 3H).

5. To a solution of S-200-N19-4_1 (1.6 g, 4.81 mmol) in DCM (20 mL) wasadded silica gel (2 g) and PCC (2.07 g, 9.62 mmol). The mixture wasstirred at 25° C. for 3 hrs. To the mixture was added PE (50 mL). Themixture was filtered though a pad of silica gel and the solid was washedwith PE/DCM (30 mL/30 mL). The mixture was filtered and the filtrate wasconcentrated under vacuum. The residue was purified by silica gelchromatography (PE/EtOAc=10/1 to 5/1) to afford S-200-N19-4_2 (1.2 g,impure) as a solid, which was re-crystallized from MeCN (10 mL) atreflux to provide S-200-N19-4_2 (1.0 g, 84.0%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.40-5.35 (m, 1H), 2.61-2.45 (m, 1H),2.30-2.10 (m, 5H), 2.00-1.75 (m, 6H), 1.70-1.10 (m, 14H), 0.90-0.75 (m,4H); 0.633 (s, 3H).

LCMS Rt=1.058 min in 2.0 min chromatography, 30-90 AB, purity 100% MSESI calcd. for C₂₂H₃₄ [M+H-H₂O]⁺ 313, found 313.

6. t-BuOK (3.51 g, 31.4 mmol) was added to a suspension of Ph₃PMeBr(11.1 g, 31.4 mmol) in THF (50 mL) under N₂ at 40° C. After stirring at25° C. for 10 min, S-200-N19-4_2 (2.6 g, 7.86 mmol) was added. Thereaction mixture was stirred at 40° C. for 1 h. The reaction wasquenched with aqueous NH₄Cl (100 mL) at 0° C., which was extracted withEtOAc (2×100 mL). The combined organic phase was washed with brine(2×100 mL), dried over Na₂SO₄, filtered and concentrated. The residuewas purified by Combi-flash (0%˜30%, EtOAc in PE) to affordS-200-N19-4_3 (2.4 g, 93%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.45-5.35 (m, 1H), 4.86-4.83 (m, 1H),8.70-4.65 (m, 1H), 2.27-2.20 (m, 1H), 2.10-1.90 (m, 4H), 1.89-1.50 (m,11H), 1.49-1.30 (m, 3H), 1.28-1.00 (m, 6H), 0.80-0.60 (m, 5H), 0.59 (s,3H). 7. 9-BBN dimer (9.27 g, 38.0 mmol) was added to a solution ofS-200-N19-4_3 (5 g, 15.2 mmol) in THF (60 mL) at 0° C. under N₂. Thesolution was stirred at 60° C. for 1 h. After cooling to 0° C., asolution of EtOH (60 ml) and NaOH (30.4 ml, 5M, 152 mmol) was added veryslowly. After addition, H₂O₂ (15.2 ml, 152 mmol, 30% in water) was addedslowly and the inner temperature was maintained below 10° C. The mixturewas stirred at 60° C. under N₂ for 1 hr. The mixture was re-cooled to30° C. Water (100 mL) was added to the solution with EtOH (100 ml). Asuspension was obtained, which was filtered and concentrated undervacuum to yield S-200-N19-4_4 (5 g, crude) as a solid.

¹H NMR (400 MHz, CDCl3) δ 5.44-5.32 (m, 1H), 3.68-3.59 (m, 1H),3.39-3.35 (m, 1H), 2.29-2.19 (m, 1H), 2.08-1.89 (m, 4H), 1.88-1.75 (m,3H), 1.62-1.60 (m, 2H), 1.56-1.39 (m, 6H), 1.36-1.24 (m, 3H), 1.23-1.11(m, 4H), 1.08-0.98 (m, 4H), 0.92-0.75 (m, 5H), 0.70 (s, 3H).

8. Dess-Martin periodinane (2.44 g, 5.76 mmol) was added to a solutionof S-200-N19-4_4 (1 g, 2.88 mmol) in DCM (150 mL) at 25° C. The reactionwas stirred at 25° C. for 1 hr. The reaction was stirred at 25° C. for30 mins. The mixture was poured into saturated Na₂S₂O₃ (100 ml) at 0°C., which was extracted with DCM (3×100 ml). The combined organic layerswere washed with saturated NaHCO₃ (100 mL×2), brine (100 mL), dried overNa₂SO₄, filtered and concentrated in vacuum to give crude product whichwas purified by a silica gel column (PE/EtOAc=10:1) to give S-500-15-2_1(800 mg, 80%) as a solid.

¹H NMR (400 MHz, CDCl3) δ 9.58-9.57 (m, 1H), 5.40-5.38 (m, 1H),2.37-2.35 (m, 1H), 2.25-2.23 (m, 1H), 2.08-1.76 (m, 7H), 1.65-1.63 (m,2H), 1.53-1.37 (m, 5H), 1.31-1.21 (m, 4H), 1.19-1.00 (m, 6H), 0.90-0.80(m, 5H), 0.73 (s, 3H).

9. A solution of 1-bromo-3-methylbutane (4 g, 26.4 mmol) in THF (27 mL)was added dropwise to a suspension of Mg (947 mg, 39.5 mmol) and I₂(33.5 mg, 0.132 mmol) in THF (3 mL) at 60° C. The mixture was stirred at60° C. for 1 hour. Freshly prepared isopentylmagnesium bromide (30 mL,0.88 M in THF, 26.4 mmol) was added to a solution of S-500-15-2_1 (800mg, 2.32 mmol) in THF (2 mL) under N₂ at 0° C. The mixture was stirredat 0° C. for 1 hour. To the mixture was added NH₄Cl (50 mL, sat. aq.).The mixture was extracted with EtOAc (2×50 mL). The combined organicphase was washed with brine (100 mL), dried over Na₂SO₄, filtered andconcentrated in vacuum to give crude product which was purified bysilica gel chromatrography (PE/EtOAc=10/1 to 5/1) to give 44 (720 mg,75%) as a solid.

¹H NMR (400 MHz, CDCl3) δ 5.40-5.38 (m, 1H), 3.63-3.61 (m, 1H),2.23-2.21 (m, 1H), 2.10-1.74 (m, 7H), 1.69-1.58 (m, 2H), 1.54-1.34 (m,8H), 1.33-1.00 (m, 11H), 0.95-0.75 (m, 14H), 0.70 (s, 3H).

LCMS Rt=1.289 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESIcalcd. For C₂₈H₄₅ [M+H-2H₂O]+ 381, found 381.

10a. To a solution of 44 (300 mg, 0.720 mmol) in THF (14 mL) was addedbenzoic acid (348 mg, 2.85 mmol) and triphenylphosphine (1.11 g, 4.27mmol) at 25° C. under N₂. After stirring at 25° C. for 20 mins, DIAD(780 mg, 3.86 mmol) was added at 0° C. under N₂. The mixture was stirredat 0° C. for 20 mins then warmed to 25° C. and stirred at 25° C. for 17hrs. Water (100 mL) was added and the mixture was extracted with EtOAc(2×100 mL). The organic phase was washed with brine (100 mL), dried overNa₂SO₄, filtered, concentrated under vacuum to give crude product (1.5g, crude) to be purified.

10b. To a solution of 44 (1.9 g, 4.55 mmol) in THF (70 mL) was addedbenzoic acid (2.19 g, 18.0 mmol) and triphenylphosphine (7.07 g, 27.0mmol) at 25° C. under N₂. After stirring at 25° C. for 20 mins, DIAD(4.93 g, 24.4 mmol) was added at 0° C. under N₂. The mixture was stirredat 0° C. for 20 mins then warmed to 25° C. and stirred at 25° C. for 17hrs. Water (250 mL) was added and the mixture extracted with EtOAc(2×250 mL). The organic phase was washed with brine (2×300 mL), driedover Na₂SO₄, filtered, concentrated in vacuum to give crude product.Combined with another batch from 300 mg of 44, to the crude product waspurified by a silica gel column (PE/EtOAc=8/1) to give S-500-15-1_1 (1.2g, impure) as an oil, which was used directly for the next step.

11. To a solution of S-500-15-1_1 (1.2 g, impure) in THF/MeOH (2 mL/2mL) was added NaOH (400 mg) and H₂O (2 mL) at 25° C. The reaction wasstirred at 50° C. for 16 h. After cooling, the reaction mixture wasdiluted with H₂O (20 mL) and extracted with EtOAc (2×30 mL). Thecombined organic layers were dried over Na₂SO₄, filtered andconcentrated under vacuum. The crude product was purified by a silicagel column (PE/EtOAc=4/1) to give the product 3 (150 mg, impure), whichwas purified by triturated with MeCN (5 mL) at 25° C. to give 3 (30 mg,pure and 100 mg, impure) as a solid.

¹H NMR (400 MHz, CDCl3) δ 5.39-5.37 (m, 1H), 3.63-3.59 (m, 1H),2.26-2.21 (m, 1H), 2.09-1.88 (m, 4H), 1.86-1.76 (m, 2H), 1.75-1.61 (m,3H), 1.54-1.32 (m, 7H), 1.32-1.08 (m, 10H), 1.07-0.96 (m, 1H), 0.95-0.74(m, 14H), 0.95-0.74 (m, 1H), 0.70 (s, 3H).

LCMS Rt=1.281 min in 2 min chromatography, 30-90 AB, purity 98%, MS ESIcalcd. For C₂₈H₄₇O [M+H-H₂O]+ 399, found 399.

Example 4: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(4)

1. t-BuLi (90.7 mL, 118 mmol, 1.3 M in n-hexane, 3.0 eq) was added to asolution of chloro(methoxymethyl)triphenylphosphorane (40.4 g, 118 mmol,3.0 eq) in THF (200 mL) at 0° C. After addition, the reaction mixturewas stirred for 1 hour at 0° C. The mixture was added to a solution ofS-500-6-29_2A (5 g, 39.6 mmol, 1.0 eq) in THF (50 mL) at 0° C. and thereaction mixture was stirred at 15° C. for 2 h. The mixture was treatedwith NH₄Cl (100 mL, 10%) and extracted with EtOAc (2×200 mL). Theorganic phase was separated and concentrated under vacuum to affordS-500-6-29_2B (18.0 g, crude).

¹H NMR (400 MHz, CDCl₃) δ 5.74 (s, 1H), 3.52 (s, 3H), 2.20-2.15 (m, 2H),1.95-1.90 (m, 2H), 1.26-1.16 (m, 4H), 0.90 (s, 6H).

2. TFA (21.4 mL, 290 mmol) was added to a stirring solution ofS-500-6-29_2B (5.6 g, impure) in DCM (25 mL) at 15° C. and stirred for1.5 h at 15° C. The reaction mixture was quenched with saturated aqueousNaHCO₃ (10 mL) and extracted with EtOAc (2×20 mL). The combined organiclayer was dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to give S-500-6-29_2C (5.0 g, crude) as an oil, which was usedfor next step without purification.

¹H NMR (400 MHz, CDCl₃) δ 9.64 (s, 1H), 2.15-2.05 (m, 1H), 1.80-1.60 (m,2H), 1.70-1.35 (m, 4H), 1.25-1.15 (m, 2H), 0.91 (s, 3H), 0.87 (s, 3H).

3. NaBH₄ (1.61 g, 42.7 mmol) was added to a solution of S-500-6-29_2C(5.0 g, 35.6 mmol) in MeOH (50 mL) at 15° C. under N₂. The mixture wasstirred at 15° C. for 1 hr. The mixture was poured into water (50 mL)and stirred for 20 minutes. The aqueous phase was extracted with EtOAc(3×50 mL). The combined organic phase was washed with saturated brine(2×50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated togive S-500-6-29_2D (5.6 g, crude) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 3.47-3.42 (m, 2H), 1.60-1.50 (m, 2H),1.42-1.30 (m, 4H), 1.25-1.0 (m, 4H), 0.91 (s, 3H), 0.87 (s, 3H).

4. TsCl (8.23 g, 43.2 mmol) was added to a solution of S-500-6-29_2D(5.6 g, 39.3 mmol) in pyridine (50 mL) at 15° C. under N₂. The mixturewas stirred at 15° C. for 16 hrs. The mixture was poured into water (50mL) and stirred for 20 minutes. The aqueous phase was extracted with DCM(3×40 mL). The combined organic phase was washed with saturated brine(2×200 mL), HCl (0.5M, 50 ml), dried over anhydrous Na₂SO₄, filtered andconcentrated to give an oil, which was re-crystallized from hexane (50mL) at 68° C. to give S-500-6-29_2E (4.2 g, 61%)} as a solid.

¹H NMR (400 MHz, CDCl₃) δ 7.80-7.76 (m, 2H), 7.35-7.25 (m, 2H),3.86-3.80 (m, 2H), 2.45 (s, 3H), 1.60-1.45 (m, 3H), 1.40-1.30 (m, 2H),1.20-1.05 (m, 4H), 0.88 (s, 3H), 0.82 (s, 3H).

5. LiBr (2.33 g, 26.9 mmol) was added to a solution of S-500-6-29_2E (2g, 6.74 mmol) in acetone (50 mL). The mixture was stirred at 65° C. for12 hrs. The mixture was quenched with water (50 mL) and extracted withMTBE (3×20 mL). The combined organic phase was washed with brine (50mL), dried over Na₂SO₄, filtered and concentrated to give S-500-6-29_2(1.3 g, crude) as a liquid. Combined with another batch from 2.2 g ofS-500-6-29_2E, the combined crude product was filtered through a smallsilicone gel and washed with PE (100 mL) and concentrated to giveS-500-6-29_2 (2.6 g, 90%) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 3.34-3.28 (m, 2H), 1.72-1.64 (m, 2H),1.60-1.48 (m, 1H), 1.42-1.35 (m, 2H), 1.28-1.18 (m, 4H), 0.91 (s, 3H),0.87 (s, 3H).

6. To a solution of Ph₃PMeBr (167 g, 470 mmol) in THF (900 mL) was addedt-BuOK (52.7 g, 470 mmol) at 25° C. The reaction mixture was heated to60° C. and stirred for 1 hour. Pregnenolone (50 g, 157 mmol) was added.The reaction mixture was stirred at 60° C. for 1 hour. Sat. NH₄Cl (900mL) was added. The mixture was extracted with EtOAc (2×1000 mL). Thecombined organic layer was washed with brine (2×2000 mL), dried overNa₂SO₄ and concentrated in vacuum to give a crude product as an oil,which was purified by column chromatography on silica gel (PE:EtOAc=20:1to 5:1) to give S-200-INT_1 (45 g, 91.2%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.40-5.30 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H),3.60-3.40 (m, 1H), 2.40-2.20 (m, 2H), 2.05-1.90 (m, 2H), 1.85-1.60 (m,9H), 1.53-1.40 (m, 5H), 1.25-0.90 (m, 9H), 0.59 (s, 3H).

7. To a solution of S-200-INT_1 (45 g, 143 mmol) in DCM (1500 mL) wasadded DMP (108 g, 257 mmol) at 20° C. The mixture was stirred at 20° C.for 2 hrs. Water (800 mL) was added and NaHCO₃ (200 g solid) was added.The mixture was filtered. The filtrate was washed with saturated Na₂S₂O₃(2×2000 mL), dried over Na₂SO₄, filtered and concentrated under vacuumto give a solution of S-200-INT_2 in DCM (100 mL), which was used in thenext step directly.

8. To a solution of BHT (191 g, 866 mmol) in toluene (500 mL) was addedAlMe₃ (2 M in toluene, 216 mL, 433 mmol) at 10° C. and stirred for 1 hr.To the mixture was added a solution of S-200-INT_2 (Theoretical Mass:44.6 g) in DCM (100 mL) at −78° C. The mixture was stirred at −78° C.for 1 hour. EtMgBr (141 mL, 426 mmol) was added at −78° C. The mixturewas stirred at −78° C. for 20 mins. Saturated citric acid (1 L) wasadded. The organic phase was separated, washed with brine (600 mL),dried over Na₂SO₄ and concentrated in vacuum to give a crude product,which was purified by column chromatography on silica gel (PE:EtOAc=50:1to 30:1) to give S-200-INT_3a (27 g, 55%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.25 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H),2.40-2.30 (m, 1H), 2.10-1.60 (m, 14H), 1.50-0.75 (m, 17H), 0.58 (s, 3H).

9. 9-BBN dimer (17.6 g, 72.5 mmol) was added to a solution ofS-200-INT_3E (5 g, 14.5 mmol) in THF (40 mL). The mixture was stirred at60° C. under N₂ for 3 hr, and a solid was formed. To the reactionmixture was added ethanol (8.33 mL, 145 mmol) and NaOH (28.9 mL, 5 M,145 mmol). The mixture turned clear. H₂O₂ (14.4 mL, 10 M, 145 mmol) wasadded dropwise at 25° C. and the inner temperature was raised to reflux(75° C.). The mixture was cooled after addition and stirred for 1 hr, asolid was formed. To the mixture was added Na₂SO₃ (20 mL, 20% aq.) at25° C. The mixture was extracted with EtOAc (2×100 mL). The combinedorganic phase was washed with brine (2×200 mL), dried over Na₂SO₄,concentrated under vacuum, and purified by silica gel column(PE/EtOAc=10/1 to 3/1) to provide S-200-INT_4E (3.5 g, 67%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.68-3.60 (m, 1H),3.41-3.32 (m, 1H), 2.40-2.32 (m, 1H), 2.03-1.93 (m, 2H), 1.92-1.65 (m,4H), 1.58-1.16 (m, 13H), 1.16-0.90 (m, 11H), 0.90-0.81 (m, 3H),0.73-0.62 (s, 3H).

10. DMP (4.66 g, 11.0 mmol) was added to a solution of S-200-INT_4E (2g, 5.54 mmol) in DCM (30 mL) at 25° C. The reaction mixture was stirredat 25° C. for 10 min. The reaction mixture was quenched with saturatedaqueous NaHCO₃ (30 mL) at 25° C. The DCM layer was separated and theaqueous phase was extracted with DCM (30 mL). The combined organic phasewas washed with saturated aqueous Na₂SO₃ (3×50 mL), brine (50 mL), driedover Na₂SO₄, filtered and concentrated under vacuum to give S-200-INT_5E(2.0 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.59-9.56 (m, 1H), 5.31-5.26 (m, 1H),2.42-2.10 (m, 2H), 2.10-1.80 (m, 4H), 1.79-1.54 (m, 7H), 1.54-1.31 (m,7H), 1.28-0.90 (m, 9H), 0.90-0.81 (m, 4H), 0.73 (s, 3H).

11. A solution of S-500-6-29_2 (2.56 g, 12.5 mmol) in THF (8 mL) wasadded dropwise to a suspension of Mg (600 mg, 25.0 mmol) and I₂ (63.4mg, 0.25 mmol) in THF (3 mL) was added at 75° C. The mixture was stirredat 75° C. for 1 hour. After cooling, a solution of S-500-6-1_1 (1 g,2.78 mmol) in THF (30 mL) was added slowly at 15° C. After addition, themixture was stirred at 15° C. for 2 hrs, quenched with saturated NH₄Cl(40 mL) and saturated citric acid (20 mL) and extracted with EtOAc (3×20mL). The combined organic phase was washed with brine (2×30 mL), driedover Na₂SO₄, filtered and concentrated and purified by combi-flash(0-15% of EtOAc in PE) to give a mixture of S-500-6-29_1 (800 mg, 60%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.19 (m, 1H), 3.88-3.71 (m, 1H),2.42-2.29 (m, 1H), 2.07-1.86 (m, 4H), 1.78-1.59 (m, 4H), 1.54-1.31 (m,13H), 1.29-1.13 (m, 8H), 1.12-0.99 (m, 8H), 0.94-0.79 (m, 13H), 0.68 (s,3H).

12. DMP (1.39 g, 3.30 mmol) was added to a solution of S-500-6-29_1 (800mg, 1.65 mmol) in DCM (30 mL). After that, the reaction mixture wasstirred at 15° C. for 10 min. The reaction mixture was quenched withsaturated NaHCO₃ aqueous (50 mL) until the pH of the aqueous layer wasabout 9. The mixture was filtered. The DCM layer was separated and theaqueous phase was extracted with DCM (20 mL). The combined organic phasewas washed with saturated Na₂S₂O₃ aqueous (3×40 mL), sat.NaHCO₃ (40 mL),brine (40 mL), dried over Na₂SO₄, filtered and concentrated to givecrude S-500-6-29_2 (800 mg, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.25 (m, 1H), 2.54-2.43 (m, 1H),2.40-2.21 (m, 3H), 2.07-1.87 (m, 3H), 1.81-1.57 (m, 7H), 1.53-1.39 (m,7H), 1.38-1.29 (m, 3H), 1.27-1.16 (m, 4H), 1.15-1.04 (m, 8H), 1.03 (s,3H), 1.00-0.92 (m, 2H), 0.91-0.80 (m, 9H), 0.69 (s, 3H).

13. NaBH₄ (2.80 g, 82.5 mmol) was added five times, every five minutes,to a solution of S-500-6-29_2 (800 mg, 1.65 mmol) in MeOH (5 mL) and THF(5 mL). The mixture was stirred at 15° C. for 30 minutes. The mixturewas quenched with saturated NH₄Cl (50 mL) and extracted with EtOAc (3×20mL). The combined organic phase was dried over Na₂SO₄, filtered,concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give49 (290 mg, 36%) and 12 (120 mg, 45%) as a solid.

49:

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.85-3.77 (m, 1H),2.40-2.32 (m, 1H), 2.07-1.87 (m, 4H), 1.76-1.69 (m, 1H), 1.66-1.55 (m,5H), 1.53-1.42 (m, 7H), 1.41-1.31 (m, 5H), 1.30-1.12 (m, 8H), 1.11-1.05(m, 3H), 1.03 (s, 3H), 1.01-0.92 (m, 2H), 0.91-0.82 (m, 12H), 0.68 (s,3H).

LCMS Rt=1.718 min in 2.0 min chromatography, 30-90AB_E, purity 98%, MSESI calcd. for C₃₃H₅₃ [M+H-2H₂O]⁺ 449, found 449.

12:

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.85-3.77 (m, 1H),2.40-2.32 (m, 1H), 2.06-1.95 (m, 3H), 1.77-1.58 (m, 7H), 1.54-1.28 (m,12H), 1.27-1.06 (m, 11H), 1.03 (s, 3H), 1.00-0.95 (m, 2H), 0.93-0.82 (m,12H), 0.69 (s, 3H).

LCMS Rt=1.708 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₃ [M+H-2H₂O]⁺ 449, found 449.

14. Pd(OH)₂ (200 mg, dry) was added to a solution of 49 (140 mg, 0.288mmol) in MeOH (30 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 59 (27 mg, 19%) and 4 (42 mg,30%) as a solid.

4:

¹H NMR (400 MHz, CDCl₃) δ 3.84-3.76 (m, 1H), 1.98-1.85 (m, 2H),1.69-1.54 (m, 9H), 1.53-1.46 (m, 3H), 1.45-1.28 (m, 9H), 1.27-1.20 (m,4H), 1.19-1.13 (m, 5H), 1.12-1.02 (m, 4H), 1.01-0.92 (m, 2H), 0.91-0.85(m, 12H), 0.82 (s, 3H), 0.70-0.61 (m, 4H).

LCMS Rt=1.799 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₅ [M+H-H₂O]⁺ 451, found 451.

Example 5: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6,6-dimethylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(5)

1. A solution of 1-bromo-3,3-dimethylbutane (3.68 g, 22.3 mmol) in THF(8 mL) was added dropwise to a suspension of Mg (1.08 g, 44.6 mmol) andI₂ (1 mg) in THF (2 mL) under N₂ at 50˜55° C. The mixture was stirred at55° C. for 1 hour. Then a solution of S-500-6-1_1 (0.8 g, 2.23 mmol) inTHF (5 mL) was added to freshly prepared (3,3-dimethylbutyl)magnesiumbromide (22.3 mmol in 10 mL of THF) at 0° C. The mixture was stirred at15° C. for 2 hrs. To the mixture was added citric acid (20 mL, 10% aq.).The mixture was extracted with EtOAc (30 mL). The organic layer wasseparated and concentrated under vacuum to give a mixture which wasseparated by flash column (0˜15% EtOAc in PE) to give 5 (580 mg, P1,58%) and 54 (50 mg, 5%, impure).

5:

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.24 (m, 1H), 3.65-3.54 (m, 1H),2.41-2.31 (m, 1H), 2.11-1.84 (m, 4H), 1.76-1.38 (m, 15H), 1.38-1.00 (m,12H), 0.93-0.80 (m, 15H), 0.70 (s, 3H).

54:

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.24 (m, 1H), 3.62-3.52 (m, 1H),2.41-2.31 (m, 1H), 2.11-1.90 (m, 3H), 1.75-1.00 (m, 28H), 1.00-0.75 (m,18H), 0.70 (s, 3H).

2. DMP (1.1 g, 2.6 mmol) and water (1 drop) were added to a solution of5 (580 mg, 1.3 mmol) in DCM (10 mL) at 20° C. The mixture was stirred at20° C. for 2 h. Saturated NaHCO₃ solution (20 mL) and Na₂S₂O₃ (20 mL,sat.) were added to the mixture. The mixture was extracted with EtOAc(50 mL). The organic layer was washed with NaHCO₃/Na₂S₂O₃ (20+20 mL,sat.) twice, dried over Na₂SO₄, filtered, concentrated under vacuum togive S-500-6-1_3 (520 mg, 90%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.38-5.18 (m, 1H), 2.62-2.22 (m, 4H),2.11-1.85 (m, 3H), 1.78-1.57 (m, 7H), 1.57-1.32 (m, 8H), 1.32-1.21 (m,2H), 1.19-1.09 (m, 5H), 1.08-1.01 (m, 4H), 1.00-0.91 (m, 1H), 0.90-0.80(m, 12H), 0.70 (s, 3H).

3. NaBH₄ (1.77 g, 46.8 mmol) was added in portions to a solution ofS-500-6-1_3 (520 mg, 1.17 mmol) in THF (5 mL) and MeOH (10 mL) at 15° C.The mixture was stirred at 15° C. for 20 min. The mixture was quenchedwith NH₄Cl (20 mL, sat. aq.) and extracted with EtOAc (50 mL). Theorganic layer was separated and concentrated in vacuum to give a mixturewhich was separated by flash column (0˜15% EtOAc in PE) to give 5 (300mg, impure) and 54 (170 mg, impure).

4. The impure 5 (300 mg, impure) was purified by flash column (0˜12%EtOAc in PE) to give a solid. The solid was dissolved in MeCN (50 mL) at60° C. and concentrated under vacuum to give 5 (270 mg, 52%) as a solid.

5:

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.24 (m, 1H), 3.67-3.54 (m, 1H),2.41-2.31 (m, 1H), 2.11-1.84 (m, 4H), 1.78-1.57 (m, 5H), 1.55-1.38 (m,12H), 1.38-1.07 (m, 7H), 1.03 (s, 3H), 0.93-0.89 (m, 12H), 0.85 (t,J=7.6 Hz, 3H), 0.70 (s, 3H).

LCMS Rt=5.587 min in 7.0 min chromatography, 30-90_AB_E, purity 96.5%,MS ESI calcd. for C₃₀H₄₉ [M+H-2H₂O]⁺ 409, found 409.

Example 6: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3R)-5-cyclopropyl-3-hydroxypentan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(6)

1. A solution of (2-bromoethyl)cyclopropane (1.8 g, 12 mmol) in THF (8mL) was added dropwise to a suspension of Mg (641 mg, 26.4 mmol) and I₂(1 mg) in THF (2 mL) under N₂ at 50˜55° C. After stirring at 55° C. for1 hour, the mixture was diluted with THF (10 mL). The Grignard solutionwas added to a solution of S-500-6-1_1 (0.8 g, 2.23 mmol) in THF (10 mL)at 0° C. After stirring at 15° C. for 4 hrs, the reaction was quenchedwith NH₄Cl (20 mL, 10% aq.) and extracted with EtOAc (30 mL). Theorganic layer was separated and concentrated under vacuum to give amixture (1 g, crude) as a solid, which was separated by flash column(0˜25% of DCM/EtOAc (1/1) in PE) to give S-500-6-20 (700 mg, 73%,impure), and S-500-6-19 (70 mg, 7%, impure) as a solid.

2. DMP (1.38 g, 3.26 mmol) and water (1 drop) were added to a solutionof S-500-6-20 (700 mg, 1.63 mmol) in DCM (10 mL) at 20° C. Afterstirring at 20° C. for 2 h, the mixture was treated with NaHCO₃ (20 mL,sat.) and Na₂S₂O₃ (20 mL, sat.) and extracted with EtOAc (50 mL). Theorganic layer was washed with saturated NaHCO₃/Na₂S₂O₃ (2×(20 mL/20mL)), dried over Na₂SO₄, filtered, and concentrated under vacuum to giveS-500-6-19_3 (700 mg, 100%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.20 (m, 1H), 2.72-2.26 (m, 4H),2.17-1.87 (m, 3H), 1.82-1.35 (m, 13H), 1.35-1.20 (m, 2H), 1.20-0.91 (m,12H), 0.85 (t, J=7.2 Hz, 3H), 0.80-0.62 (m, 4H), 0.53-0.33 (m, 2H),0.12-0.00 (m, 2H).

3. NaBH₄ (2.46 g, 65.1 mmol) was added in portions to a solution ofS-500-6-1_3 (700 mg, 1.63 mmol) in THF (5 mL) and MeOH (5 mL) at 15° C.After stirring at 15° C. for 20 mins, the mixture was quenched withNH₄Cl (20 mL, sat. aq.) and extracted with EtOAc (50 mL). The organiclayer was separated and concentrated under vacuum to give 760 mg mixtureas a solid, which was separated by flash column (0˜35% of DCM/EtOAc(1/1) in PE) to give 69 (330 mg, 47%) and 6 (250 mg, 35%, impure) as asolid. The impure 6 (250 mg) was further separated by flash column(0˜35% of DCM/EtOAc (1/1) in PE) to give 6 (170 mg, 23%) as a solid.

6:

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.24 (m, 1H), 3.77-3.66 (m, 1H),2.41-2.31 (m, 1H), 2.09-1.91 (m, 3H), 1.79-1.59 (m, 6H), 1.55-1.21 (m,14H), 1.21-1.06 (m, 4H), 1.03 (s, 3H), 1.00-0.95 (m, 1H), 0.93 (d, J=6.8Hz, 3H) 0.85 (t, J=7.6 Hz, 3H), 0.70 (s, 3H), 0.68-0.62 (m, 1H),0.49-0.38 (m, 2H), 0.11-0.02 (m, 2H).

LCMS Rt=1.380 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₉H₄₇O [M+H-H₂O]⁺ 411, found 411.

Example 7: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((1R,2S)-1-hydroxy-1-(pyridin-3-yl)propan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(7)

1. To a solution of BHT (416 g, 1.88 mol) in toluene (1500 mL) under N₂at 0° C. was added trimethylaluminum (2 M in toluene, 469 mL, 939 mmol)dropwise. The mixture was stirred at 0° C. for 30 mins and used directlyas a solution of MAD (0.47 M in toluene) without further purification.To a solution of MAD (0.47 M in toluene, 2.01 L, 945 mmol) under N₂ at−70° C. was added a solution of N-005_1 (100 g, 315 mmol) in toluene(800 mL) dropwise. The mixture was stirred at −70° C. for 30 mins. Tothe above mixture was added EtMgBr (3 M in ethyl ether, 315 mL, 945mmol) dropwise. The resulting mixture was stirred at −70° C. for 1 hr.The reaction mixture was poured to ice-cooled aqueous citric acid (1000mL), extracted with EtOAc (2×600 mL). The combined organic layer waswashed with brine (500 mL), dried over anhydrous sodium sulfate,filtered and concentrated. The residue was purified by silica gelchromatography (0-20% of EtOAc in PE) to give 85 g of N-005_2 as a solid(78% yield).

¹H NMR (400 MHz, CDCl₃) δ 2.55-2.46 (m, 1H), 2.19-2.12 (m, 1H),2.11-2.09 (m, 3H), 2.08-1.96 (m, 1H), 1.71-1.48 (m, 10H), 1.47-1.31 (m,5H), 1.30-1.09 (m, 7H), 1.06-0.94 (m, 2H), 0.92-0.87 (m, 3H), 0.86-0.79(m, 3H), 0.75-0.64 (m, 1H), 0.60 (s, 3H).

2. To a suspension of MePPh₃Br (174 g, 0.49 mol) in THF (1000 mL) wasadded t-BuOK (54.9 g, 0.49 mol) at 15° C. under N₂. After stirring at50° C. for 30 mins, a solution of N-005_2 (85 g, 245 mmol) in THF (800mL) was added in portions below 65° C. The mixture was stirred at 50° C.for 1 hr, quenched with NH₄Cl (1000 mL), extracted with EtOAc (2×900mL). The organic layer was separated, concentrated in vacuum to give acrude product which was triturated from MeOH/water (1.5 L, 1:1) at 50°C. The mixture was filtered after cooled and the filter cake was washedwith MeOH/water (2×500 mL, 1:1), concentrated in vacuum to give N-005_3(75 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.85-4.82 (m, 1H), 4.71-4.68 (m, 1H),2.06-1.94 (m, 1H), 1.86-1.78 (m, 1H), 1.76-1.71 (m, 4H), 1.70-1.62 (m,4H), 1.61-1.48 (m, 6H), 1.47-1.30 (m, 3H), 1.29-1.05 (m, 8H), 1.04-0.92(m, 1H), 0.91-0.82 (m, 6H), 0.76-0.63 (m, 1H), 0.56 (s, 3H).

3. To a solution of N-005_3 (75 g, 217 mmol) in THF (1800 mL) was added9-BBN dimer (105 g, 434 mmol) under N₂. The mixture was stirred at 60°C. for 3 hrs. To the reaction mixture was added ethanol (124 mL, 2.17mol) and NaOH aqueous (434 mL, 5 M, 2.17 mmol) in portions. Then H₂O₂(217 mL, 10 M, 2.17 mol) was added dropwise at 0° C. The mixture waswarmed to 65° C. and stirred for 1 hr and diluted with water (1.5 L).The reaction mixture was extracted with EtOAc (2×800 mL). The combinedorganic layer was added saturated aqueous Na₂S₂O₃ (600 mL) and stirredfor 1 hour. The reaction was checked by potassium iodide-starch testpaper to confirm excess H₂O₂ was destroyed. Then the organic phase waswashed with saturated brine (2×500 mL), dried over anhydrous Na₂SO₄,filtered and concentrated in vacuum to give N-005_4 (78 g, crude) as asolid. The crude N-005_4 (78 g, impure) was triturated fromMeOH/H2O=10/1 at 15° C. to give N-005_4 (70 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.68-3.60 (m, 1H), 3.41-3.32 (m, 1H),1.99-1.92 (m, 1H), 1.88-1.75 (m, 1H), 1.69-1.45 (m, 10H), 1.44-1.29 (m,4H), 1.28-1.15 (m, 6H), 1.14-0.91 (m, 8H), 0.90-0.79 (m, 7H), 0.67 (s,3H).

4. To a solution of N-005_4 (70 g, 193 mmol) in DCM (800 mL) was addedDMP (122 g, 289 mmol). After that, the reaction was stirred at 15° C.for 30 mins. The reaction mixture was added saturated aqueous NaHCO₃(500 mL) solution and stirred at 15° C. for 20 mins. Saturated aqueousNa₂S₂O₃ (600 mL) was added and the mixture was stirred at 15° C. foranother 1 hr. The reaction was checked by potassium iodide-starch testpaper to confirm excess DMP was destroyed. The aqueous phase wasextracted with DCM (2×400 mL). The combined organic layer was washedwith aqueous saturated NaHCO₃ (400 mL) solution and brine (400 mL),dried over Na₂SO₄, filtered and concentrated in vacuum to give N-005_5(70 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.58-9.55 (m, 1H), 2.39-2.30 (m, 1H),1.95-1.78 (m, 2H), 1.69-1.42 (m, 10H), 1.41-1.30 (m, 4H), 1.29-1.14 (m,5H), 1.13-0.95 (m, 6H), 0.94-0.86 (m, 4H), 0.85-0.81 (m, 3H), 0.69 (m,4H).

5. i-PrMgCl (2.49 mL, 4.98 mmol, 2M in ether) was added to a solution of3-bromopyridine (875 mg, 5.54 mmol) in THF (5 mL) dropwise. Afterstirring at 25° C. for 1 h, a solution of N-8-7_1 (200 mg, 0.554 mmol)in THF (5 mL) was added. After stirring at 25° C. for 16 hrs, thereaction mixture was quenched with NH₄Cl (50 mL, 10% aq.) and extractedwith EtOAc (2×20 mL). The combined organic phase was dried over Na₂SO₄,filtered, concentrated and purified by flash column (0˜50% of EtOAc inDCM) to give N-8-19_1 (100 mg, 41%) as a solid.

6. N-8-19_1 (100 mg, 0.227 mmol) was separated by SFC (column: AD (250mm*30 mm, 5 um), gradient: 50-50% B (A=0.05% NH₃/H₂O, B=MeOH), flowrate: 80 mL/min) to give 7 (Peak 1, 57 mg, 57%) and 89 (Peak 2, 8 mg,8%) as a solid.

SFC Peak 1: Rt=1.798 min and Peak 2 Rt=1.985 min in 3 minchromatography, AD-H_3UM_4_5_40_4ML (“Chiralpak AD-3 50*4.6 mm I.D., 3um Mobile phase: A: CO₂ B: iso-propanol (0.05% DEA) Gradient: from 5% to40% of B in 1.4 min and hold 40% for 1.05 min, then 5% of B for 0.35 minFlow rate: 4 mL/min Column temp.: 40° C.″).

7:

¹H NMR (400 MHz, CDCl₃) δ 8.56-8.52 (m, 1H), 8.49-8.45 (m, 1H),7.68-7.62 (m, 1H), 7.29-7.24 (m, 1H), 5.01-4.95 (m, 1H), 2.11-2.01 (m,1H), 1.96-1.89 (m, 1H), 1.83-1.76 (m, 1H), 1.73-1.63 (m, 4H), 1.59-1.47(m, 6H), 1.43-1.29 (m, 4H), 1.27-1.20 (m, 4H), 1.19-1.06 (m, 4H),1.03-0.92 (m, 1H), 0.91-0.85 (m, 4H), 0.83 (s, 3H), 0.77-0.73 (m, 3H),0.70-0.64 (m, 4H).

LCMS Rt=1.017 min in 2.0 min chromatography, 10-80AB_E, purity 100%, MSESI calcd. for C₂₉ H₄₆NO₂ [M+H]⁺ 440, found 440.

SFC Rt=1.780 min in 3 min chromatography, AD-H_3UM_4_5_40_4ML, 100% de.

Example 8: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(8)

1. To a solution of BHT (1.97 kg, 8.94 mol) in toluene (1 L) was addedAlMe₃ (2.14 L, 2.0 M in toluene, 4.28 mol) drop-wise blew 25° C. underN₂ atmosphere. The resulting mixture was stirred at 25° C. for 1 hour.S-200-INT_2 (794 g, 85% percent weight, 2.16 mol) in DCM (3 L) was addedat −70° C. The mixture was stirred at −70° C. for 1 hour. MeMgBr (862mL, 3.0 M in diethyl ether, 2.59 mol) was added at −70° C. The reactionmixture was stirred at −70° C. for 10 mins. The mixture was quenched bysaturated critic acid (3 L), extracted with EtOAc (2×2 L). The combinedorganic phase was washed with brine (2 L), dried over Na₂SO₄, filteredand concentrated under vacuum to give a residue, which was trituratedfrom MeCN (3 L) at 25° C. to give S-200-INT_3 (340 g, 43%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.34-5.26 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H),2.50-2.35 (m, 1H), 2.07-1.94 (m, 3H), 1.91-1.84 (m, 1H), 1.83-1.63 (m,8H), 1.58-1.33 (m, 6H), 1.27-1.13 (m, 3H), 1.12 (s, 3H), 1.10-1.05 (m,1H), 1.02 (s, 3H), 1.00-0.92 (m, 1H), 0.58 (s, 3H).

2. To a mixture of S-200-INT_3 (149 g, 453 mmol) and 9-BBN dimer (127 g,520 mmol) was added THF (1 L) at 15° C. under N₂. The reaction mixturewas stirred at 60° C. for 1 hour. The mixture was cooled to 15° C. EtOH(208 g, 4.53 mol) was added at 15° C. NaOH aqueous (906 mL, 5 M, 4.53mol) was added drop-wise at 15° C. H₂O₂ (514 g, 30%, 4.53 mol) was addeddropwise at 15° C. The obtained mixture was stirred at 60° C. for 1hour. A solid was produced. The solid was washed with ethanol (200 mL)to give a solid, which was triturated with EtOH (2.3 L) at reflux andwater (2.5 L) at 80° C. successively to give 15-3b (131 g, 84%) as asolid. The filtrate from ethanol was concentrated under vacuum to give15-3b (30 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ5.35-5.24 (m, 1H), 3.67-3.61 (m, 1H), 3.42-3.33(m, 1H), 2.50-2.35 (m, 1H), 2.07-1.92 (m, 3H), 1.88-1.65 (m, 3H),1.60-1.38 (m, 9H), 1.37-1.26 (m, 1H), 1.26-1.12 (m, 4H), 1.11 (s, 3H),1.08 (s, 1H), 1.05 (d, J=6.8 Hz, 3H), 1.01 (s, 3H), 1.00-0.91 (m, 1H),0.70 (s, 3H).

3. DMP (2.44 g, 5.76 mmol) was added to a solution of 15-3b (1 g, 2.88mmol) in DCM (10 mL). After that, the reaction was stirred at 25° C. for10 min. The reaction mixture was quenched by adding aqueous saturatedNaHCO₃ solution (20 mL) and aqueous saturated Na₂S₂O₃ (20 mL) solution,extracted with DCM (2×50 mL). The combined organic layer was washed withaqueous saturated NaHCO₃ (3×50 mL) solution and brine (50 mL), driedover Na₂SO₄, filtered and concentrated under vacuum to give S-500-2-9_1(1 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.57 (brs, 1H), 5.35-5.25 (m, 1H), 2.50-2.30(m, 2H), 2.05-1.95 (m, 3H), 1.95-1.80 (m, 1H), 1.75-1.65 (m, 1H),1.65-1.60 (m, 3H), 1.55-1.50 (m, 2H), 1.50-1.40 (m, 2H), 1.40-1.30 (m,1H), 1.25-1.20 (m, 2H), 1.20-1.15 (m, 2H), 1.15-1.10 (m, 6H), 1.05-0.95(m, 5H), 0.90-0.70 (m, 1H), 0.68 (s, 3H).

4. A mixture of magnesium (641 mg, 26.4 mmol) and I₂ (33.5 mg, 0.132mmol) was stirred at 60° C. and a solution of isopentylmagnesium bromide(2 g, 13.2 mmol) in THF (20 mL) was added dropwise under N₂. After that,the reaction mixture was stirred at 60° C. for 1 hr. The reactionmixture was used directly as isopentylmagnesium bromide solution withoutany purification. The Grignard solution was added to a solution ofS-500-2-9_1 (1 g, 2.90 mmol) in THF (10 mL) at 0° C. under N₂. Afterthat, the reaction mixture was stirred at 25° C. for 1 hr. The reactionmixture was added saturated aqueous NH₄Cl (50 mL) solution, extractedwith EtOAc (2×50 mL), washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated in vacuum to give a crude product. The crudeproduct was purified by silica gel column (EtOAc/PE=¼) to give impureS-500-2-9-1A (560 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.28-5.25 (m, 1H), 3.90-3.80 (m, 0.25H),3.68-3.58 (m, 0.75H), 2.48-2.36 (m, 1H), 2.05-1.95 (m, 3H), 1.95-1.80(m, 1H), 1.80-1.75 (m, 1H), 1.75-1.52 (m, 6H) 1.52-1.42 (m, 6H),1.42-1.32 (m, 3H), 1.32-1.22 (m, 3H), 1.22-1.12 (m, 3H), 1.12-1.02 (m,2H), 1.01 (s, 3H), 1.00-0.92 (m, 1H), 0.92-0.85 (m, 9H), 0.85-0.77 (m,1H), 0.69 (s, 3H).

5. S-500-2-9-1A (560 mg) was purified by SFC (Column: Chiralcel OD-3150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA)Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5%of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.) to giveimpure 30 (160 mg) as a solid and 75 (265 mg, 47%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.30 (m, 1H), 3.70-3.60 (m, 1H),2.50-2.40 (m, 1H), 2.05-1.90 (m, 4H), 1.85-1.75 (m, 2H), 1.75-1.60 (m,1H), 1.55-1.45 (m, 8H), 1.45-1.25 (m, 8H), 1.25-1.10 (m, 4H), 1.10-1.05(m, 2H), 1.02 (s, 3H), 0.99-0.91 (m, 3H), 0.91-0.89 (m, 4H), 0.88 (s,3H), 0.69 (s, 3H).

LCMS Rt=1.162 min in 1.5 min chromatography, 5-95 AB, purity 99%, MS ESIcalcd. for C₂₈H₄₅ [M+H-2H₂O]⁺ 381, found 381.

6. Dry Pd(OH)₂/C (100 mg) was added to a solution of 75 (230 mg, 0.551mmol) in THF (5 mL) and MeOH (5 mL). The reaction mixture was stirred at50° C. for 24 h under H₂ and 50 Psi. After that, HNMR showed thereaction was completed. The reaction mixture was filtered with filterpaper and concentrated in vacuum to give impure product. The impureproduct was re-crystallized with MeCN (3 mL) to give 8 (68 mg, 30%) asan off-white solid.

¹H NMR (400 MHz, CDCl₃) δ 3.65-3.55 (m, 1H), 2.00-1.80 (m, 2H),1.76-1.60 (m, 3H), 1.55-1.48 (m, 3H), 1.48-1.38 (m, 4H), 1.38-1.26 (m,7H), 1.26-1.23 (m, 4H), 1.23-1.06 (m, 5H), 1.06-1.02 (m, 3H), 1.02-095(m, 1H), 0.95-0.85 (m, 10H), 0.81 (s, 3H), 0.70-0.60 (m, 4H).

LCMS Rt=1.171 min in 1.5 min chromatography, 5-95 AB, purity 100%.

MS MS ESI calcd. for C₂₈H₄₇ [M+H-2H₂O]⁺ 383, found 383.

Example 9: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-(methoxymethyl)-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(9)

1. Pd/C (5 g, <1% water) was added to a solution of N-004-022_1 (50 g,151 mmol) in MeOH (100 mL) and THF (100 mL). Then the solution washydrogenated under 30 psi of hydrogen at 25° C. for 48 hrs. The mixturewas filtered through a pad of celite and the filtrate was concentratedin vacuum to give N-004-022_2 (50 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.65-3.55 (m, 1H), 3.40-3.3 (m, 1H), 2.80-2.60(m, 1H), 2.40-2.30 (m, 1H), 2.25-2.10 (m, 1H), 2.10-1.95 (m, 3H),1.80-1.65 (m, 3H), 1.65-1.53 (m, 1H), 1.53-1.40 (m, 4H), 1.40-1.01 (m,17H), 0.70 (s, 3H).

2. A stirred solution of trimethylsulfoxonium iodide (19.8 g, 90.2 mmol)and t-BuOK (10.1 g, 90.2 mmol) in DMSO (200 mL) was heated at 60° C. for1 hr under N₂. N-004-022_2 (15 g, 45.1 mmol) was added to the reactionmixture and stirred at 60° C. for 10 mins. The reaction was treated withwater (1000 mL), extracted with EtOAc (2×500 mL). The combined organicphase was washed with water (2×500 mL), brine (300 mL), dried overanhydrous Na₂SO₄, filtered, concentrated under vacuum to affordN-004-022_3 (15.5 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.18-4.08 (m, 1H), 3.67-3.60 (m, 1H),3.40-3.30 (m, 1H), 2.70-2.50 (m, 3H), 2.40-2.30 (m, 1H), 2.01-1.50 (m,14H), 1.40-0.65 (m, 14H), 0.68 (s, 3H).

3. MeONa (12.0 g, 223 mmol) was added to a solution of N-004-022_3 (15.5g, 44.7 mmol) in MeOH (500 mL) at 25° C. under N₂. The mixture wasstirred at 70° C. reflux for 16 hrs under N₂. The reaction was treatedwith water (500 mL). The aqueous phase was extracted with DCM (2×300mL). The combined organic phase was washed with saturated brine (2×300mL), dried over anhydrous Na₂SO₄, filtered and concentrate to giveN-004-022_4 (15 g, crude) as a solid. The crude N-004-022_4 (15 g) waspurified by silica gel chromatography (PE/EtOAc=10/1 to 5/1) to affordN-004-022_4 (7.4 g, 50%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.76-3.73 (m, 1H), 3.64-3.60 (m, 1H),3.40-3.33 (m, 4H), 3.22-3.16 (m, 2H), 2.01-1.69 (m, 6H), 1.62-1.51 (m,4H), 1.44-1.31 (m, 13H), 1.10-0.99 (m, 5H), 0.97 (s, 3H), 0.67 (s, 3H).

4. DMP (1.56 g, 3.69 mmol) was added to a solution of N-004-022_4 (1.4g, 3.69 mmol) in DCM (15 mL). After that, the reaction mixture wasstirred at 25° C. for 10 min. The reaction mixture was quenched withsaturated NaHCO₃ aqueous (20 mL) until pH=9. The mixture was filtered.The DCM layer was separated and the aqueous phase was extracted with DCM(20 mL). The combined organic phase was washed with saturated Na₂S₂O₃aqueous solution (3×10 mL), sat. NaHCO₃ solution (10 mL), brine (20 mL),dried over Na₂SO₄, filtered and concentrated. The residue was trituratedwith MeCN (10 mL) to give N-004-022_5 (700, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.56-9.58 (m, 1H), 3.39 (s, 3H), 3.24-3.18 (m,2H), 2.40-2.31 (m, 1H), 2.01-1.50 (m, 11H), 1.47-1.01 (m, 16H), 0.97 (s,3H), 0.70 (s, 3H).

5. To a solution of N-004-022_5 (200 mg, 0.531 mmol), CsF (40.2 mg,0.265 mmol) in THF (5 mL) was added TMSCF₃ (187 mg, 1.32 mmol) under N₂at 0° C. The mixture was stirred at 25° C. for 1 hrs. To the mixture wasadded TBAF.3H₂O (836 mg, 2.65 mmol). After stirring at 25° C. for 2 hrs,the mixture was quenched 50% NH₄Cl (20 mL) and extracted with EtOAc(2×10 mL). The combined organic phase was washed with brine (20 mL),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. Theresidue was purified by silica gel chromatography (100-200 mesh silicagel, PE/EA=10/1) to afford 9 (56 mg, 24%) and 71 (30 mg, impure) as awhite solid.

9:

¹H NMR (400 MHz, CDCl₃) δ 4.05-3.95 (m, 1H), 3.39 (s, 3H), 3.24-3.18 (m,2H), 2.00-1.83 (m, 5H), 1.77-1.68 (m, 2H), 1.64-1.47 (m, 8H), 1.43-1.35(m, 5H), 1.31-1.08 (m, 6H), 1.06-1.00 (m, 3H), 0.97 (s, 3H), 0.70 (s,3H).

LCMS Rt=1.156 min in 2 min chromatography, 30-90AB_2 min.1 cm, purity100%, MS ESI calcd. for C₂₅H₄₁F₃O₃ [M+Na]⁺ 469, found 469.

Example 10: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((1S,2S)-1-cyclopropyl-1-hydroxypropan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(10)

1. N-8-7_1 (500 mg, 1.38 mmol) in THF (5 mL) was added to a solution ofcyclopropylmagnesium bromide (1 g, 13.7 mL, 0.5M in THF) in THF (5 mL)at 0° C. and stirred for 4 hrs at 25° C. The mixture was added NH₄Cl (20mL, 10% aq.) and extracted with EtOAc (2×30 mL). The organic layer wasseparated and concentrated in vacuum to give a residue. The residue waspurified by silica gel chromatography eluted with PE/EtOAc=1/1 to affordN-8-13_1 (140 mg, 25%) as a solid.

LCMS Rt=1.192 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 99%,MS ESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

2. DMP (294 mg, 0.694 mmol) was added to a solution of N-8-13_1 (140 mg,0.347 mmol) in DCM (5 mL). After that, the reaction mixture was stirredat 25° C. for 1 h. The reaction mixture was quenched with saturatedNaHCO₃ aqueous (50 mL) until the pH of the aqueous layer became about 9.The mixture was filtered. The DCM layer was separated and the aqueousphase was extracted with DCM (100 mL). The combined organic phase waswashed with saturated Na₂S₂O₃ aqueous solution (3×100 mL), sat. NaHCO₃(100 mL), brine (40 mL), dried over Na₂SO₄, filtered and concentrated togive N-8-13_2 (140 mg, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.65-2.55 (m, 1H), 1.95-1.90 (m, 2H),1.50-1.15 (m, 19H), 1.14-0.95 (m, 7H), 0.94-0.80 (m, 12H), 0.69 (s, 3H).

3. NaBH₄ (1.18 g, 17.4 mmol) was added five times, every five minutes,to a solution of N-8-13_2 (140 mg, 0.347 mmol) in MeOH (1 mL) and THF (1mL). The mixture was stirred at 15° C. for 30 minutes. The mixture wasquenched with sat. NH₄Cl (50 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was dried over Na₂SO₄, filtered, concentrated andpurified by combi-flash (25% of EtOAc in PE) to give 10 (26 mg, 19%) asa solid and 90 (12 mg, 9%) as a solid.

10:

¹H NMR (400 MHz, CDCl₃) δ 2.85-2.80 (m, 1H), 2.00-1.95 (m, 1H),1.90-1.80 (m, 1H), 1.55-1.10 (m, 16H), 1.09-0.80 (m, 17H), 0.70-0.60 (m,5H), 0.58-0.43 (m, 3H), 0.32-0.34 (m, 1H), 0.13-0.06 (m, 1H).

LCMS Rt=3.840 min in 7.0 min chromatography, 30-90AB_7MIN_E, purity 97%,MS ESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

HPLC Rt=13.470 min in 30 min chromatography, 70-90AB_1_30MIN.M, purity97%.

Example 11: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(11)

1. TMSCF₃ (493 mg, 3.47 mmol) was added to a solution of S-500-6-1_1(500 mg, 1.39 mmol) and CsF (105 mg, 695 umol) in THF (5 mL) at 0° C.The mixture was stirred at 25° C. for 1 hr and treated with TBAF.3H₂O(1.09 g, 3.47 mmol). The mixture was stirred at 25° C. for 2 hrs,quenched with water (100 mL) and extracted with EtOAc (2×50 mL). Thecombined organic phase was washed with brine (100 mL), dried overanhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue waspurified by silica gel chromatography (100-200 mesh silica gel,PE/EA=10/1) to afford S-500-6-1_2 (400 mg, 67%) as a solid.

¹H NMR (400 MHz, CDCl3) δ 5.33-5.24 (m, 1H), 4.06-4.00 (m, 1H),2.38-2.35 (m, 1H), 2.08-1.82 (m, 6H), 1.77-1.69 (m, 1H), 1.62-1.20 (m,13H), 1.16-1.00 (m, 8H), 0.99-0.92 (m, 1H), 0.87-0.83 (m, 4H), 0.74-0.64(m, 3H).

2. S-500-6-1_2 (350 mg) was purified by SFC (Column: AD (250 mm*30 mm, 5um), Condition: 0.1% NH₃.H₂O ETOH, Gradient: from 35% to 35%, Flow Rate(ml/min): 60 mL/min, 25° C.) to afford 81 (Peak 1, 130 mg, 37%) and 62(Peak 2, 180 mg, 52%) as a solid.

81:

¹H NMR (400 MHz, CDCl3) δ 5.34-5.24 (m, 1H), 4.09-4.00 (m, 1H),2.43-2.33 (m, 1H), 2.14 (d, J=4 Hz, 1H), 2.07-1.80 (m, 5H), 1.77-1.55(m, 5H), 1.53-1.30 (m, 7H), 1.28-1.00 (m, 11H), 1.00-0.91 (m, 1H), 0.85(t, J=8 Hz, 3H), 0.70 (s, 3H).

LCMS Rt=1.220 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₃₈F₃O [M+H-H₂O]⁺ 411, found 411.

SFC Peak 1: Rt=4.561 min in 10 min chromatography,AD_3_EtOH_DEA_5_40_25ML (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 umMobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% ofB in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate:2.5 mL/min Column temp.: 35° C.″), 100% de.

3. Pd(OH)₂ (0.2 g, <1% water) was added to a solution of 81 (110 mg,0.256 mmol) in MeOH (2 mL) and THF (1 mL). The solution was hydrogenatedunder 50 psi of hydrogen at 50° C. for 48 hrs. Then the mixture wasfiltered through a pad of celite and the filtrate was concentrated undervacuum. The residue was purified by flash column (PE/EtOAc=10/1 to 5/1)to give 56 (38 mg, 35%) and 11 (42 mg, 38%) as a solid.

11:

¹H NMR (400 MHz, CDCl₃) δ 4.09-3.99 (m, 1H), 2.11 (d, J=6.0 Hz, 1H),1.99-1.80 (m, 3H), 1.70-1.55 (m, 6H), 1.53-1.30 (m, 8H), 1.27-0.96 (m,11H), 0.95-0.81 (m, 7H), 0.70-0.61 (m, 4H).

LCMS Rt=1.247 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₄₀F₃O [M+H-H₂O]⁺ 413, found 413.

Example 12: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3R)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(12)

1. NaBH₄ (2.80 g, 82.5 mmol) was added five times, every five minutes,to a solution of S-500-6-29_2 (800 mg, 1.65 mmol) in MeOH (5 mL) and THF(5 mL). The mixture was stirred at 15° C. for 30 minutes. The mixturewas quenched with sat. NH₄Cl (50 mL) and extracted with EtOAc (3×20 mL).The combined organic phase was dried over Na₂SO₄, filtered, concentratedand purified by combi-flash (0-15% of EtOAc in PE) to give S-500-6-30(290 mg, 36%) and 12 (120 mg, 45%) as a solid.

12:

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.85-3.77 (m, 1H),2.40-2.32 (m, 1H), 2.06-1.95 (m, 3H), 1.77-1.58 (m, 7H), 1.54-1.28 (m,12H), 1.27-1.06 (m, 11H), 1.03 (s, 3H), 1.00-0.95 (m, 2H), 0.93-0.82 (m,12H), 0.69 (s, 3H).

LCMS Rt=1.708 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₃ [M+H-2H₂O]⁺ 449, found 449.

Example 13: Synthesis of(1S,3R,4S)-4-((3S,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1-phenylpentane-1,3-diol(13)

1. To a solution of N-4-1_4 (27 g, 85.8 mmol) in THF (200 mL) was addedCsF (25.9 g, 171 mmol) and TMSCF₃ (24.3 g, 171 mmol). The mixture wasstirred at 10° C. for 1 hr. To the mixture was added water (10 mL) andTBAF.3H₂O (30 g). The mixture was stirred at 30° C. for another 2 hrs.The mixture was concentrated under vacuum. The residue was dissolved inEtOAc (500 mL), washed with water (2×500 mL), dried over Na₂SO₄,filtered, concentrated under vacuum and purified by flash column(DCM/EtOAc (1:1) in PE, 0˜10%) to give N-4-1_5 (27 g, 82%) and N-4-1_5A(3.5 g, 11%) as a solid.

N-4-1_5:

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.12-1.94 (m, 3H),1.89-1.78 (m, 2H), 1.75 (s, 3H), 1.72-1.60 (m, 5H), 1.58-1.48 (m, 2H),1.45-1.09 (m, 10H), 1.01-0.89 (m, 1H), 0.85 (s, 3H), 0.78-0.68 (m, 1H),0.56 (s, 3H).

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.09-1.99 (m, 1H),1.89-1.78 (m, 2H), 1.75 (s, 3H), 1.72-1.52 (m, 9H), 1.45-1.06 (m, 10H),1.00-1.81 (m, 2H), 0.79 (s, 3H), 0.56 (s, 3H).

2. To a solution of N-4-1_5 (23 g, 59.8 mmol) in THF (250 mL) was added9-BBN dimer (29 g, 119 mmol) was stirred at 40° C. under N₂ for 16 hrs.To the reaction mixture was added ethanol (34.3 mL, 598 mmol) and NaOH(119 mL, 5 M, 598 mmol). The mixture turned clear. H₂O₂ (59.8 mL, 10 M,598 mmol) was added dropwise at 25° C. and the inner temperature wasraised to reflux (70° C.). The mixture was cooled to 30° C. after theaddition. To the mixture was added Na₂SO₃ (100 mL, 20% aq.). The organiclayer was separated and poured into water (800 mL). A solid was formed.The mixture was filtered and the solid was washed with water, dried invacuum and triturated with MeCN (250 mL) to give a solid. The solid wastriturated form MeOH/water (250 mL/12.5 mL) at 60° C. and filtered aftercooled to 15° C. The solid was dried in vacuum to give N-4-1_6 (16.4 g,68%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.69-3.60 (m, 1H), 3.39-3.29 (m, 1H),2.09-2.01 (m, 1H), 1.99-1.92 (m, 1H), 1.87-1.75 (m, 2H), 1.72-1.43 (m,7H), 1.42-1.07 (m, 11H), 1.03 (d, J=6.8 Hz, 3H), 1.01-0.86 (m, 3H), 0.85(s, 3H), 0.73-0.69 (m, 1H), 0.67 (s, 3H).

3. To a suspension of N-4-1_6 (5 g, 12.4 mmol) in DCM (200 mL) was addedwater (223 mg, 12.4 mmol) and DMP (10.5 g, 24.8 mmol). The mixture wasstirred at 15° C. for 15 mins. The mixture was washed withNaHCO₃/Na₂S₂O₃ (200 mL/200 mL, sat.) twice, dried over Na₂SO₄, filteredand concentrated in vacuum to give N-4-1_7 (4.5 g, 90%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.60-9.51 (m, 1H), 2.40-2.30 (m, 1H),2.12-1.78 (m, 5H), 1.75-1.59 (m, 4H), 1.57-1.15 (m, 11H), 1.14-0.84 (m,8H), 0.78-0.63 (m, 5H).

4. MeLi (7.75 mL, 1.6 M, 12.4 mmol) was added to a solution of N-4-1_7(1 g, 2.49 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 15°C. for 1 h. To the mixture was added NH₄Cl (10%, 20 mL). The mixture wasextracted with EtOAc (2×30 mL). The combined organic layer was driedover Na₂SO₄, filtered and concentrated in vacuum to give a mixture (1 g)as agum. The mixture N-3-2_1 (1 g) was purified by flash column (0˜15%EtOAc in PE) to give 91 (450 mg) and 22 (460 mg) and 130 mg of amixture. The 91 (450 mg) was re-crystallized form MeCN (10 mL) to give91 (50 mg) as a solid, 22 (460 mg) was re-crystallized twice form MeCN(10 mL) to give 22 (50 mg) as a solid.

91:

¹H NMR (400 MHz, CDCl₃) δ 3.98-3.88 (m, 1H), 2.11-2.02 (m, 1H), 2.00 (s,1H), 1.98-1.88 (m, 2H), 1.85-1.79 (m, 1H), 1.73-1.58 (m, 4H), 1.52-1.20(m, 11H), 1.19-1.11 (m, 4H), 1.10-1.00 (m, 3H), 0.97-0.89 (m, 4H), 0.85(s, 3H), 0.75-0.68 (m, 1H), 0.66 (s, 3H).

LCMS Rt=1.155 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

HPLC Rt=5.23 min in 10.0 min chromatography, 30-90_AB_E, purity 98.88%,d.e. 100%.

22:

¹H NMR (400 MHz, CDCl₃) δ 3.97-3.82 (m, 1H), 2.10-1.92 (m, 3H),1.85-1.78 (m, 1H), 1.77-1.60 (m, 5H), 1.59-1.06 (m, 13H), 1.05-0.81 (m,12H), 0.74-0.62 (m, 4H).

LCMS Rt=1.136 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

HPLC Rt=5.05 min in 10.0 min chromatography, 30-90_AB_E, purity 100%,d.e. 100%.

5. To a solution of N-3-2_1 (0.88 g, 2.11 mmol) in DCM (20 mL) was addedwater (2 drops) and DMP (1.78 g, 4.22 mmol). The mixture was stirred at25° C. for 30 min. The mixture was washed with NaHCO₃/Na₂S₂O₃ (30 mL/30mL, sat.) twice, dried over Na₂SO₄, filtered and concentrated in vacuumto give N-3-2_2 (0.85 g, 97%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.53-2.42 (m, 1H), 2.13-2.00 (m, 4H),1.97-1.78 (m, 2H), 1.75-1.45 (m, 9H), 1.43-1.13 (m, 9H), 1.11 (d, J=8.4Hz, 3H), 1.07-1.00 (m, 1H), 0.98-0.88 (m, 1H), 0.85 (s, 3H), 0.78-0.68(m, 1H), 0.67 (s, 3H).

6. To a solution of N-3-2_2 (0.85 g, 2.05 mmol) in DCM (5 mL) was addedimidazole (279 mg, 4.10 mmol) and TMSCl (333 mg, 3.07 mmol) at 0° C. Themixture was stirred at 0° C. for 0.5 h. The mixture was added quenchedby NaHCO₃ (20 mL, sat) and extracted with PE (15 mL). The organic layerwas separated, dried over Na₂SO₄, filtered and concentrated in vacuum togive N-3-2_2A (0.98 g, 98%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.53-2.42 (m, 1H), 2.13-2.03 (m, 4H),1.97-1.78 (m, 2H), 1.75-1.31 (m, 11H), 1.31-1.00 (m, 10H), 1.00-0.88 (m,1H), 0.83 (s, 3H), 0.75-0.61 (m, 4H), 0.15 (s, 9H).

7. BuLi (0.384 mL, 2.5 M, 0.615 mmol) was added to i-Pr₂NH (62.2 mg,0.615 mmol) in THF (0.5 mL) at −70° C. and stirred at 0° C. for 10 min.N-3-2_2A (0.2 g, 0.41 mmol) in THF (1 mL) was added at −70° C. andstirred at −70° C. for 30 min. A solution of benzaldehyde (91.3 mg,0.861 mmol) in THF (0.5 mL) was added at −70° C. and stirred at −70° C.for 15 min. NH₄Cl (1 mL, sat., aq.) was added to the mixture andextracted with EtOAc (10 mL). The organic layer was separated, driedover Na₂SO₄, filtered and concentrated in vacuum to give N-3-2_3C (250mg, crude) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 7.46-7.30 (m, 5H), 5.70-5.55 (m, 1H),3.60-3.25 (m, 1H), 2.90-2.70 (m, 2H), 2.55-2.41 (m, 1H), 2.16-2.00 (m,2H), 1.96-1.75 (m, 3H), 1.50-1.15 (m, 9H), 1.13-1.05 (m, 4H), 1.05-0.88(m, 4H), 0.87-0.80 (m, 5H), 0.73-0.62 (m, 5H), 0.15 (s, 9H).

8. LiAlH₄ (159 mg, 4.20 mmol) was added to a solution of N-3-2_3C (250mg, 0.421 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 0° C.for 5 min. Water (0.16 mL), NaOH (0.16 mL, 15% aq.), and water (0.48 mL)were added to the mixture in the order written here. The mixture wasfiltered and the solid was washed with THF (30 mL). The combinedfiltrate was concentrated in vacuum to give N-3-2_4C (250 mg, 100%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 7.42-7.28 (m, 5H), 5.70-5.30 (m, 1H),4.15-3.65 (m, 1H), 2.18-1.55 (m, 9H), 1.53-1.00 (m, 15H), 1.00-0.75 (m,9H), 0.75-0.50 (m, 4H), 0.15 (s, 9H).

9. TBAF (219 mg, 0.84 mmol) was added to a solution of N-3-2_4C (250 mg,0.42 mmol) in THF (2 mL). The mixture was stirred at 25° C. for 3 h. Themixture was concentrated in vacuum. The residue was dissolved in EtOAc(10 mL) and washed with water (3×10 mL), purified by flash column(10˜25% EtOAc in PE) to give N-3-2_5C (150 mg, 68%) as a solid.

10. The mixture N-3-2_5C (150 mg) was separated by SFC (Instrument:MG-II; Column: IC (250 mm*30 mm, 10 um); Condition: 0.1% NH₃H₂O MeOH;Begin B: 30%; End B: 30%; FlowRate (mL/min): 60; Injections: 300) togive impure 79 (35 mg, impure), mixture of 31 and 25 (55 mg) and 13 (28mg, impure).

The impure 79 (35 mg) was purified by flash column (10˜30% EtOAc in PE),the eluent was concentrated in vacuum. The residue was dissolved inMeCN/water (20 mL, 4:1) and concentrated in vacuum to give 79 (12 mg) asa solid.

25 and 13 (55 mg) was separated by SFC (Instrument: MG-II; Column: AD(250 mm*30 mm, 5 um); Condition: 0.1% NH₃H₂O MeOH; Begin B: 35%; End B:35%; FlowRate (mL/min): 60; Injections: 70). Each of the eluent wasrespectively concentrated in vacuum, dissolved in MeCN/water (20 mL,4:1) and concentrated in vacuum to give 25 (28 mg) and 13 (7 mg) both asa solid.

The impure 31 (28 mg) was purified by SFC (Instrument: SFC 17; Column:AS (250 mm*30 mm, 5 um); Condition: 0.1% NH₃H₂O EtOH; Begin B: 30%; EndB: 30%; FlowRate (mL/min): 50; Injections: 60) to give a solid. Theresidue was dissolved in MeCN/water (20 mL, 4:1) and concentrated invacuum to give 31 (9 mg) as a solid.

SFC of four isomer: Peak 1: Rt=1.501 min, Peak 2: Rt=1.730 min and Peak3: Rt=1.943 min in 10 min chromatography, IC-3_MeOH(DEA)_40_2.5ML(“Column: ChiralPak IC-3 150×4.6 mm I.D., 3 um; Gradient:40% of Methanol(0.05% DEA) in CO₂; Flow rate: 2.5 mL/min; Column temperature: 40° C.”.

SFC of 25 and 13: Peak 1: Rt=4.411 min and Peak 2: Rt=4.920 min in 8 minchromatography, AD_MEOH(DEA)_5_40_2,8ML_8MIN (“Column: Chiralpak AD-3100×4.6 mm I.D., 3 um; Mobile phase: A: CO2 B: Methanol (0.05% DEA);Gradient: from 5% to 40% of B in 4.5 min and hold 40% for 2.5 min, then5% of B for 1 min; Flow rate: 2.8 mL/min; Column temperature: 40° C.”.

79:

¹H NMR (400 MHz, CDCl₃) δ 7.43-7.28 (m, 5H), 5.05-4.94 (m, 1H),4.04-3.91 (m, 1H), 2.51 (brs, 1H), 2.07-1.78 (m, 6H), 1.70-1.61 (m, 4H),1.51-1.41 (m, 3H), 1.39-1.12 (m, 11H), 1.05-0.98 (m, 2H), 0.91-0.81 (m,7H), 0.71-0.60 (m, 4H).

LCMS Rt=1.298 min in 2 min chromatography, 10-80AB_2MIN_E, purity 96.7%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.483 min in 10 min chromatography, IC-3_MeOH(DEA)_40_2.5ML, 100%de.

25:

¹H NMR (400 MHz, CDCl₃) δ 7.42-7.28 (m, 5H), 4.97-4.81 (m, 1H),4.12-3.92 (m, 1H), 3.23 (brs, 1H), 2.69 (brs, 1H), 2.10-1.88 (m, 3H),1.82-1.62 (m, 7H), 1.48-1.18 (m, 10H), 1.10-0.88 (m, 8H), 0.87-0.78 (m,4H), 0.70-0.58 (m, 4H).

LCMS Rt=1.319 min in 2 min chromatography, 10-80AB_2MIN_E, purity 97.0%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.718 min in 5 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.26% de.

SFC Rt=4.367 min in 8 min chromatography, AD_MEOH(DEA)_5_40_2,8ML_8MIN,100% de.

31:

¹H NMR (400 MHz, CDCl₃) δ 7.45-7.28 (m, 5H), 5.02-4.81 (m, 1H),4.18-3.98 (m, 1H), 3.35 (brs, 1H), 2.47 (brs, 1H), 2.15-1.72 (m, 8H),1.53-1.31 (m, 8H), 1.30-1.03 (m, 8H), 0.99-0.89 (m, 4H), 0.89-0.78 (m,4H), 0.75-0.60 (m, 4H).

LCMS Rt=1.327 min in 2 min chromatography, 10-80AB_2MIN_E, purity 100%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.929 min in 10 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.4% de.

13:

¹H NMR (400 MHz, CDCl₃) δ 7.40-7.28 (m, 5H), 5.12-5.07 (m, 1H),3.95-3.88 (m, 1H), 2.76 (brs, 1H), 2.08-1.78 (m, 6H), 1.75-1.60 (m, 5H),1.51-1.38 (m, 4H), 1.36-1.09 (m, 9H), 1.00-0.89 (m, 6H), 0.83 (s, 3H),0.71-0.64 (m, 1H), 0.63 (s, 3H).

LCMS Rt=1.309 min in 2 min chromatography, 10-80AB_2MIN_E, purity 100%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.683 min in 5 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.94% de.

SFC Rt=4.785 min in 8 min chromatography, AD_MEOH(DEA)_5_40_2,8ML_8MIN,94.03% de.

Example 14: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R,E)-3-hydroxy-5-phenylpent-4-en-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(14)

1. BuLi (0.384 mL, 2.5 M, 0.615 mmol) was added to i-Pr₂NH (62.2 mg,0.615 mmol) in THF (0.5 mL) at −70° C. and stirred at 0° C. for 10 min.N-3-2_2A (0.2 g, 0.41 mmol) in THF (1 mL) was added at −70° C. andstirred at −70° C. for 1 h. A solution of benzaldehyde (91.3 mg, 0.861mmol) in THF (0.5 mL) was added at −70° C. and stirred at 20° C. for 4h. NH₄Cl (1 mL, sat., aq.) was added to the mixture and extracted withEtOAc (10 mL). The organic layer was separated, dried over Na₂SO₄,filtered and concentrated in vacuum, purified by flash column (0˜10%EtOAc in PE) to give N-3-2_3D (150 mg, 64%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 7.64-7.55 (m, 3H), 7.43-7.39 (m, 3H), 6.79 (d,J=16.0 Hz, 1H), 2.86-2.73 (m, 1H), 2.15-2.08 (m, 1H), 2.00-1.90 (m, 1H),1.88-1.80 (m, 1H), 1.72-1.59 (m, 5H), 1.53-1.22 (m, 9H), 1.21-1.03 (m,7H), 0.99-0.88 (m, 1H), 0.84 (s, 3H), 0.75-0.61 (m, 4H), 0.15 (s, 9H).

2. TBAF (135 mg, 0.52 mmol) was added to a solution of N-3-2_3D (150 mg,0.26 mmol) in THF (1 mL). The mixture was stirred at 20° C. for 20 h. Tothe mixture was added EtOAc (5 mL). The mixture was washed with water(2×5 mL), brine (5 mL), concentrated in vacuum to give N-003-005_1 (140mg, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 7.64-7.53 (m, 3H), 7.43-7.35 (m, 3H), 6.79 (d,J=16.0 Hz, 1H), 2.88-2.73 (m, 1H), 2.13-1.90 (m, 3H), 1.88-1.78 (m, 1H),1.77-1.90 (m, 5H), 1.60-1.22 (m, 8H), 1.21-0.88 (m, 9H), 0.86 (s, 3H),0.75-0.61 (m, 4H).

3. NaBH₄ (419 mg, 11.1 mmol) was added in portions to a solution ofN-003-005_1 (140 mg, 0.278 mmol) in THF (2 mL) and MeOH (1 mL) at 20° C.The mixture was stirred at 20° C. for 10 min. The reaction was quenchedwith water (20 mL) and NH₄Cl (20 mL, sat.). The mixture was extractedwith EtOAc (50 mL). The organic layer was concentrated in vacuum andpurified by prep-TLC (PE/EtOAc=4/1) to give N-003-005 (50 mg, impure)and N-003-006 (50 mg) both as a solid. 14 (50 mg) was dissolved in MeCN(20 mL) and concentrated in vacuum to give 14 (29 mg) as a solid.

14:

¹H NMR (400 MHz, CDCl₃) δ 7.44-7.38 (m, 2H), 7.37-7.29 (m, 2H),7.25-7.18 (m, 1H), 6.59 (d, J=16.0 Hz, 1H), 6.24 (dd, J=7.2, 16.0 Hz,1H), 4.40-4.30 (m, 1H), 2.08-1.92 (m, 3H), 1.89-1.77 (m, 3H), 1.68-1.60(m, 3H), 1.50-1.08 (m, 13H), 1.03-0.82 (m, 9H), 0.72-0.62 (m, 4H).

LCMS Rt=1.236 min in 2 min chromatography, 30-90AB_2MIN_E, purity 99.0%,MS ESI calcd. for C₃₁H₄₂F₃O [M+H-H₂O]⁺ 487, found 487.

HPLC Rt=5.89 min in 8 min chromatography, 30-90_AB_1.2 ml, 98.1% d.e.(220 nm)

Example 15: Synthesis(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6,6-dimethylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(15)

1. Pd(OH)₂/C (100 mg) was added to a solution of 44 (100 mg) in MeOH/THF(2 mL/2 mL) was added. The mixture was stirred at 50° C. under H₂ (50psi) for 20 h. The mixture was filtered. The filtered was concentratedto give 100 mg of a solid. NMR showed 9% 54 was remained. The impuresample was hydrogenated at the same condition for another 3 times. Themixture was filtered. The filtrate was concentrated and separated byflash column (0˜15% EtOAc in PE) to give 5 (7 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.48 (m, 1H), 2.00-1.55 (m, 9H),1.50-1.22 (m, 15H), 1.19-1.03 (m, 8H), 0.96 (s, 3H), 0.91-0.81 (m, 15H),0.67 (s, 3H).

LCMS Rt=1.492 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₃₀H₅₁ [M+H-2H₂O]⁺ 411, found 411.

Example 16: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(16)

1. Pd(OH)₂ (0.2 g, <1% water) was added to a solution of 62 (160 mg,0.373 mmol) in MeOH (2 mL) and THF (1 mL). The solution was hydrogenatedunder 50 psi of hydrogen at 50° C. for 16 hrs. Then the mixture wasfiltered through a pad of celite and the filtrate was concentrated invacuum. The residue was purified by flash column (PE/EtOAc=10/1 to 5/1)to give 44 (27 mg, 17%) and 16 (117 mg, 73%) as solids.

16:

¹H NMR (400 MHz, CDCl3) δ 4.04-3.96 (m, 1H), 1.98-1.83 (m, 4H),1.69-1.59 (m, 3H), 1.56-1.20 (m, 13H), 1.17-0.95 (m, 8H), 0.91-0.83 (m,8H), 0.70-0.62 (m, 4H).

LCMS Rt=1.240 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₄₀F₃O [M+H-H₂O]⁺ 413, found 413.

Example 17: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6,6-dimethylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(17)

1. Pd(OH)₂/C (100 mg) was added to a solution of 5 (250 mg) in MeOH/THF(2 mL/2 mL). The mixture was stirred at 50° C. under H₂ (50 psi) for 20hrs. The mixture was filtered. The filtered was concentrated to give 250mg of a solid. NMR showed 70% of 5 remained. The impure sample washydrogenated at the same condition for another 3 times. The mixture wasfiltered. The filtrate was concentrated and separated by flash column(0˜15% EtOAc in PE) to give 17 (3 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.63-3.50 (m, 1H), 1.98-1.55 (m, 8H),1.49-1.37 (m, 8H), 1.35-1.21 (m, 8H), 1.19-1.01 (m, 8H), 0.97 (s, 3H),0.91-0.82 (m, 15H), 0.66 (s, 3H).

LCMS Rt=1.529 min in 2.0 min chromatography, 30-90_AB_E, purity 95.6%,MS ESI calcd. for C₃₀H₅₁ [M+H-2H₂O]⁺ 411, found 411.

Example 18: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-(tetrahydro-2H-pyran-4-yl)butan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(18)

1. To suspension of Me₃SI (8.44 g, 41.4 mmol) in anhydrous THF (50 mL)at 20° C. under nitrogen was added t-BuOK (4.64 g, 41.4 mmol). Themixture was stirred at 20° C. for 1 hr, and N-005_5 (5 g, 13.8 mmol) wasadded. The resulting mixture was warmed to 45° C. and stirred for 4 hrs.The reaction mixture was cooled to room temperature, quenched with water(50 mL) and extracted with EtOAc (2×50 mL). The combined organic layerwas dried over anhydrous sodium sulfate, filtered and concentrated. Theresidue was purified by column chromatography on silica gel (0˜10% EtOAcin PE) to give N-005_001 (2.7 g, 52%) as a solid.

LCMS Rt=1.324 min in 2.0 min chromatography, 30-90AB_E, purity 92%, MSESI calcd. for C₂₅H₄₁O [M+H-H₂O]⁺ 357, found 357.

2. A solution of 4-chlorotetrahydro-2H-pyran (1 g, 8.29 mmol) inanhydrous THF (8 mL) was added dropwise to a mixture of Mg (401 mg, 16.5mmol) and I₂ (105 mg, 0.414 mmol) in anhydrous THF 2 mL) under N₂ at 60°C. The mixture was stirred at 60° C. for 10 min. The temperature rose to66° C. The reaction mixture was stirred for additional 30 min, cooled toroom temperature which was used directly as a solution of(tetrahydro-2H-pyran-4-yl)magnesium chloride (0.83 M in THF).

3. A solution of (tetrahydro-2H-pyran-4-yl)-magnesium chloride (0.83 Min THF, 6.38 mL, 5.30 mmol) was added dropwise to a suspension ofN-005_001 (400 mg, 1.06 mmol) and CuI (20.1 mg, 0.106 mmol) in anhydrousTHF (10 mL) under nitrogen at 20° C. The mixture was stirred at 20° C.for 18 hrs. The reaction mixture was quenched with water (10 mL) andsaturated NH₄Cl (10 mL), extracted with EtOAc (2×15 mL). The combinedorganic layer was washed with brine (15 mL), dried over anhydrous sodiumsulfate, filtered and concentrated to give N-6-9/10 (760 mg, crude) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 4.01-3.92 (m, 3H), 3.83-3.62 (m, 1H),3.42-3.32 (m, 3H), 1.97-1.87 (m, 1H), 1.68-1.58 (m, 7H), 1.57-1.45 (m,6H), 1.43-1.29 (m, 8H), 1.24-0.95 (m, 10H), 0.91-0.79 (m, 9H), 0.73-0.56(m, 4H).

LCMS Rt=1.332 min in 2.0 min chromatography, 10-80AB, purity 93%, MS ESIcalcd. for C₃₀H₅₂NaO₃ [M+Na]⁺ 483, found 483.

4. BzCl (691 mg, 4.92 mmol) and DMAP (20 mg, 0.164 mmol) were added to asolution of N-6-9/10 (760 mg, 1.64 mmol) in pyridine (10 mL). Themixture was stirred at 20° C. for 2 hrs. The reaction mixture wasquenched with water (15 mL), extracted with EtOAc (2×20 mL). Thecombined organic layer was washed with 10% aqueous HCl (2×20 mL),saturated NaHCO₃ (40 mL) and brine (20 mL), dried over anhydrous sodiumsulfate, filtered and concentrated. The residue was purified by columnchromatography on silica gel (0˜10% of EtOAc in PE) to give N-6-9_1 (400mg, 43%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 8.08-8.01 (m, 1H), 8.08-8.01 (m, 1H),7.61-7.42 (m, 3H), 5.42-5.29 (m, 1H), 3.99-3.85 (m, 2H), 3.41-3.24 (m,2H), 2.06-1.65 (m, 5H), 1.65-1.57 (m, 5H), 1.54-1.42 (m, 6H), 1.42-1.14(m, 11H), 1.14-0.90 (m, 8H), 0.89-0.77 (m, 7H), 0.69-0.51 (m, 4H).N-6-9_1 (400 mg, 0.708 mmol) was separated and purified by SFC (column:C2 250 mm*30 mm, 10 um, gradient: 45-45% B (A=0.1% NH₃/H₂O, B=MeOH),flow rate: 60 mL/min) to give N-6-9_2 (peak 1, Rt=3.926 min, 80 mg, 20%)as a solid and N-6-10_1 (peak 2, Rt=4.893 min, 180 mg, 45%) as a solid.

N-6-9_2:

¹H NMR (400 MHz, CDCl₃) δ 8.06-8.00 (m, 2H), 7.60-7.53 (m, 1H),7.49-7.42 (m, 2H), 5.35-5.28 (m, 1H), 3.98-3.91 (m, 2H), 3.41-3.31 (m,2H), 1.88-1.67 (m, 5H), 1.66-1.57 (m, 4H), 1.54-1.36 (m, 10H), 1.35-1.16(m, 8H), 1.08-0.88 (m, 8H), 0.88-0.82 (m, 4H), 0.80 (s, 3H), 0.64 (s,3H), 0.61-0.54 (m, 1H).

LCMS Rt=1.540 min in 2.0 min chromatography, 30-90AB, purity 96%, MS ESIcalcd. for C₃₀H₄₉O [M-BzOH-H₂O+H]⁺ 425, found 425.

SFC Rt=3.789 min in 8 min chromatography, Column: Lux Cellulose-2150×4.6 mm I.D., 3 μm; Mobile phase: 40% of Methanol (0.05% DEA) in CO₂;Flow rate: 2.5 mL/min; Column temperature: 40° C., 97% de.

N-6-10_1:

¹H NMR (400 MHz, CDCl₃) δ 8.06-8.01 (m, 2H), 7.61-7.53 (m, 1H),7.49-7.42 (m, 2H), 5.41-5.33 (m, 1H), 3.98-3.86 (m, 2H), 3.40-3.27 (m,2H), 2.05-1.91 (m, 2H), 1.84-1.72 (m, 2H), 1.66-1.59 (m, 3H), 1.55-1.38(m, 9H), 1.37-1.16 (m, 11H), 1.13-1.00 (m, 6H), 1.00-0.90 (m, 2H),0.89-0.79 (m, 7H), 0.67 (s, 3H), 0.63-0.54 (m, 1H).

LCMS Rt=1.507 min in 2.0 min chromatography, 30-90AB, purity 97%, MS ESIcalcd. for C₃₀H₄₉O [M-BzOH-H₂O+H]⁺ 425, found 425.

SFC Rt=4.699 min in 8 min chromatography, Column: Lux Cellulose-2150×4.6 mm I.D., 3 μm; Mobile phase: 40% of Methanol (0.05% DEA) in CO₂;Flow rate: 2.5 mL/min; Column temperature: 400° C., 97% de.

5. Water (1 mL) and KOH (78.5 mg, 1.40 mmol) were added to a solution ofN-6-9_2 (80 mg, 0.141 mmol) in THF (2 mL) and methanol (1 mL). Themixture was stirred at 50° C. for 18 hrs. The reaction mixture wascooled, diluted with water (5 mL), acidified with 10% HCl (0.2 mL) andextracted with EtOAc (3×5 mL). The combined organic layer was dried overanhydrous sodium sulfate, filtered and concentrated. The residue waspurified by column chromatography on silica gel (10˜30% of EtOAc in PE)to give 18 (13 mg, 20%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.05-3.89 (m, 3H), 3.45-3.34 (m, 2H),1.94-1.87 (m, 1H), 1.86-1.76 (m, 1H), 1.72-1.59 (m, 6H), 1.54-1.45 (m,4H), 1.44-1.28 (m, 9H), 1.28-1.15 (m, 7H), 1.13-0.92 (m, 5H), 0.91-0.85(m, 4H), 0.84-0.79 (m, 6H), 0.70-0.62 (m, 4H).

LCMS Rt=1.213 min in 2.0 min chromatography, 30-90AB, purity 100%.

MS MS ESI calcd. for C₃₀H₄₉O [M-2H₂O+H]⁺ 425, found 425.

Example 19: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((1S,2S)-1-hydroxy-1-phenylpropan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(19)

1. A solution N-8-7_1 (300 mg, 0.832 mmol) in THF (5 mL) was added to asolution of PhMgBr (1.38 mL, 3 M in ether, 4.15 mmol) in THF (10 mL) at0° C., then the reaction mixture was stirred at 0° C. for 3 hours. Next,the reaction mixture was stirred at 25° C. for 5 hrs. The reactionmixture was quenched by water (10 mL) at 0° C. The solution was filteredand the filter cake was washed with EtOAc (10 mL). The aqueous phase wasextracted with EtOAc (3×15 mL). The combined organic phase was washedwith saturated brine (2×10 mL), dried over anhydrous Na₂SO₄, filteredand concentrated. The residue was purified by silica gel chromatography(PE/EtOAc=20/1 to 1/1) to afford crude product (200 mg) as a solid. Thecrude product was purified by SFC (column: AD (250 mm*30 mm, 5 um)),gradient: 25-25% B (A=0.1% NH3/H2O, B=EtOH), flow rate: 60 mL/min) togive 55 (Peak2, 55 mg, 15%) and 19 (Peak1, 21 mg, 6%) as a solid.

19:

¹H NMR (400 MHz, CDCl₃) δ 7.40-7.20 (m, 5H), 4.85-4.80 (m, 1H),2.10-1.60 (m, 5H), 1.55-1.05 (m, 17H), 0.95-0.75 (m, 14H), 0.71 (s, 3H),0.60-0.50 (m, 1H).

LCMS Rt=1.208 min in 2.0 min chromatography, 30-90AB_2 min., purity100%, MS ESI calcd. For C₃₀H₄₃ [M-2H₂O+H]⁺ 403, found 403.

SFC Rt=1.047 min in 3 min chromatography, OJ_3_EtOH_DEA_5_40_25ML, 100%de.

Example 20: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(20)

1. Pd(OH)₂ (150 mg, dry) was added to a solution of 12 (100 mg, 0.206mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 87 (12 mg, 12%) and 20 (11mg, 11%) as a solid.

20:

¹H NMR (400 MHz, CDCl₃) δ 3.83-3.75 (m, 1H), 1.99-1.92 (m, 1H),1.71-1.57 (m, 8H), 1.52-1.43 (m, 3H), 1.41-1.29 (m, 8H), 1.27-1.13 (m,11H), 1.12-1.04 (m, 4H), 1.03-0.94 (m, 3H), 0.91-0.86 (m, 12H), 0.82 (s,3H), 0.68-0.59 (m, 4H).

LCMS Rt=1.748 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₅ [M+H-H₂O]⁺ 451, found 451.

Example 21: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxypentan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(21)

1. EtMgBr (0.553 mL, 3 M in ether, 1.66 mmol) was added dropwise to asolution of N-8_1 (250 mg, 0.8320 mmol) in THF (3 mL) at 25° C. underN₂. The mixture was stirred at 25° C. for 1 hr, quenched with sat. NH₄Cl(10 mL) and extracted with EtOAc (3×15 mL). The organic layers werewashed with brine (20 mL), dried over Na₂SO₄, filtered, concentratedunder vacuum to give a crude product N-8-24_1, which was purified byflash column (0˜15% of EtOAc in PE) to give 21 (130 mg, impure) as asolid. The impure N-8-24 (130 mg, 0.3327 mmol) was re-crystallized fromMeCN (3 mL) at 85° C. to give pure 21 (111 mg, 86%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.62-3.50 (m, 1H), 2.02-1.81 (m, 2H),1.72-1.59 (m, 3H), 1.56-1.46 (m, 4H), 1.45-1.17 (m, 12H), 1.16-1.00 (m,5H), 0.99-0.85 (m, 11H), 0.84-0.78 (m, 4H), 0.66 (s, 4H).

HPLC Rt=5.73 min in 10 min chromatography, 30-90_AB_1.2 mL_E, purity100%.

MS MS ESI calcd. for C₂₆H₄₃ [M+H-2H₂O]⁺ 355, found 355.

Example 22: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxybutan-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(22)

1. MeLi (7.75 mL, 1.6 M, 12.4 mmol) was added to a solution of N-4-1_7(1 g, 2.49 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 15°C. for 1 hr. To the mixture was added NH₄Cl (10%, 20 mL). The mixturewas extracted with EtOAc (2×30 mL). The combined organic layer was driedover Na₂SO₄, filtered and concentrated in vacuum to give a mixture (1 g)as a gum. The mixture (1 g) was purified by flash column (0˜15% EtOAc inPE) to give 91 (450 mg) and 22 (460 mg) and 130 mg of a mixture. 91 (450mg) was re-crystallized from MeCN (10 mL) to give 91 (50 mg) as a solid.22 (460 mg) was re-crystallized twice form MeCN (10 mL) to give 22 (50mg) as a solid.

22:

¹H NMR (400 MHz, CDCl₃) δ 3.97-3.82 (m, 1H), 2.10-1.92 (m, 3H),1.85-1.78 (m, 1H), 1.77-1.60 (m, 5H), 1.59-1.06 (m, 13H), 1.05-0.81 (m,12H), 0.74-0.62 (m, 4H).

LCMS Rt=1.136 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

HPLC Rt=5.05 min in 10.0 min chromatography, 30-90_AB_E, purity 100%,d.e. 100%.

Example 23: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxyhexan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(23)

1. Propylmagnesium bromide (3.34 mL, 6.69 mmol, 2M in THF) was slowlyadded to a solution of S-500-6-1_1 (800 mg, 2.23 mmol) in THF (30 mL) at0° C. After addition, the mixture was stirred at 15° C. for 1 hr. Themixture was quenched with sat.NH₄Cl (40 mL) and extracted with EtOAc(3×20 mL). The combined organic phase was washed with brine (2×30 mL),dried over Na₂SO₄, filtered and concentrated and purified by combi-flash(0-15% of EtOAc in PE) to give 72 (500 mg, 56%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.72-3.64 (m, 1H),2.41-2.31 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.69 (m, 1H), 1.62-1.54 (m,3H), 1.52-1.38 (m, 9H), 1.37-1.16 (m, 6H), 1.15-1.01 (m, 7H), 0.99-0.88(m, 7H), 0.87-0.82 (m, 3H), 0.68 (s, 3H).

LCMS Rt=4.979 min in 7.0 min chromatography, 30-90AB_E, purity 98.8%, MSESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

2. Pd(OH)₂ (300 mg, dry) was added to a solution of 72 (150 mg, 0.372mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 23 (9 mg, 6%) and 38 (43 mg,29%) as solids.

23:

¹H NMR (400 MHz, CDCl₃) δ 3.71-3.62 (m, 1H), 2.01-1.83 (m, 3H),1.82-1.72 (m, 1H), 1.69-1.57 (m, 3H), 1.51-1.37 (m, 9H), 1.36-1.22 (m,9H), 1.20-1.00 (m, 8H), 0.97 (s, 3H), 0.94-0.87 (m, 8H), 0.66 (s, 3H).

LCMS Rt=1.440 min in 2.0 min chromatography, 30-90AB_E, purity 98.8%, MSESI calcd. for C₂₇H₄₅ [M+H-2H₂O]⁺ 369, found 369.

Example 24: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(24)

1. A solution of 1-bromo-3-methylbutane (11.7 g, 78 mmol) in THF (8 mL)was added dropwise to a suspension of Mg (4.35 g, 179 mmol) and 12 (20mg) in THF (2 mL) at 60° C. The mixture was stirred at 60° C. for 1 hr.The mixture was diluted with THF (10 mL) and used directly. Freshlyprepared isopentylmagnesium bromide (19.5 mL, 3.9 M in THF, 76 mmol) wasadded to a solution of S-200-INT_5E (1.0 g, 2.78 mmol) in THF (5 mL)under N₂ at 0° C. The mixture was stirred at 0° C. for 1 hr. NH₄Cl (20mL, sat. aq.) was added to the mixture. The mixture was extracted withEtOAc (2×30 mL). The combined organic phase was washed with brine (100mL), dried over Na₂SO₄, concentrated in vacuum, purified by silica gel(PE/EtOAc=20/1 to 10/1), and re-crystallized from CH₃CN (10 mL) to 77(255 mg, 21%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.26 (m, 1H), 3.66-3.59 (m, 1H),2.42-2.32 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.58 (m, 4H), 1.55-1.38 (m,10H), 1.38-1.19 (m, 5H), 1.19-1.00 (m, 8H), 1.00-0.81 (m, 13H), 0.69 (s,3H).

LCMS Rt=1.306 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. for C₂₉H₄₉O [M+H-H₂O]⁺ 413, found 413.

2. Benzoic acid (508 mg, 4.16 mmol) and triphenylphosphine (1.63 g, 6.24mmol) were added to a solution of 77 (900 mg, 2.08 mmol) in THF (30 mL)at 25° C. under N₂. The mixture was stirred at 25° C. for 20 mins. DIAD(1.26 g, 6.24 mmol) was added at 0° C. under N₂. The mixture was stirredat 0° C. for 20 mins, warmed to 25° C. and stirred at 25° C. for 16 hrs.The reaction mixture was quenched with water (60 mL) and extracted withMTBE (3×30 mL). The combined organic phase was washed with brine (60mL), dried over Na₂SO₄, filtered, concentrated under vacuum to give acrude product, which was purified by flash column (0-10% EtOAc in PE) togive impure product S-500-2-15_1 (900 mg) as an oil, which was useddirectly for the next step.

3. NaOH solution (974 mg in 6 mL H₂O, 16.8 mmol) was added to a solutionof S-500-2-15_1 (900 mg, 1.68 mmol) in THF (10 mL) and MeOH (5 mL). Themixture was heated at 50° C. for 16 hrs. The reaction mixture wasquenched with sat. NH₄Cl (60 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was washed with brine (60 mL), dried over Na₂SO₄,filtered concentrated, and purified by combi-flash (0-15% of EtOAc inPE) to give 210 mg of a solid, which was purified by SFC (column: AD(250 mm*30 mm, 5 um), gradient: 35-35% B (A=0.1% NH₃/H₂O, B=MeOH), flowrate: 80 mL/min) to give 52 (150 mg, 68%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.30-5.26 (m, 1H), 3.64-3.58 (m, 1H),2.40-2.30 (m, 1H), 2.02-1.92 (m, 3H), 1.80-1.58 (m, 7H), 1.56-1.31 (m,9H), 1.30-1.05 (m, 8H), 1.03 (s, 3H), 1.02-0.96 (m, 2H), 0.95-0.86 (m,9H), 0.85-0.80 (m, 3H), 0.69 (s, 3H).

LCMS t_(R)=1.335 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₉H₄₇ [M+H-2H₂O]⁺ 395, found 395.

4. Pd(OH)₂ (200 mg) was added to a solution of 52 (50 mg, 0.116 mmol) inMeOH (10 mL). The mixture was stirred at 50° C. under H₂ (50 Psi). Themixture was filtered, concentrated and purified by combi-flash (0-10% ofEtOAc in PE) to give 24 (15 mg, 30%) as a solid and 96 (1.2 mg, 3%) as asolid.

24:

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.52 (m, 1H), 2.02-1.91 (m, 1H),1.74-1.57 (m, 7H), 1.52-1.44 (m, 2H), 1.43-1.29 (m, 7H), 1.28-1.04 (m,11H), 1.03-0.94 (m, 3H), 0.94-0.85 (m, 13H), 0.82 (s, 3H), 0.71-0.60 (m,4H).

LCMS t_(R)=1.342 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₉H₄₉ [M+H-2H₂O]⁺ 397, found 397.

Example 25: Synthesis of(1S,3S,4S)-4-((3S,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1-phenylpentane-1,3-diol(25)

The preparation of 25 can be found in Example 13.

25:

¹H NMR (400 MHz, CDCl₃) δ 7.42-7.28 (m, 5H), 4.97-4.81 (m, 1H),4.12-3.92 (m, 1H), 3.23 (brs, 1H), 2.69 (brs, 1H), 2.10-1.88 (m, 3H),1.82-1.62 (m, 7H), 1.48-1.18 (m, 10H), 1.10-0.88 (m, 8H), 0.87-0.78 (m,4H), 0.70-0.58 (m, 4H).

LCMS Rt=1.319 min in 2 min chromatography, 10-80AB_2MIN_E, purity 97.0%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.718 min in 5 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.26% de.

SFC Rt=4.367 min in 8 min chromatography, AD_MEOH(DEA)_5_40_2,8ML_8MIN,100% de.

Example 26: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((S)-3-hydroxy-3-methylbutan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(26)

1. MeMgBr (0.83 mL, 2.49 mmol, 3M in ether) was added dropwise to asolution of N-8-7_1 (300 mg, 0.832 mmol) in THF (20 mL) at 0° C. underN₂. After stirring at 20° C. for 30 minutes, the reaction mixture wasquenched with sat. NH₄Cl (50 mL) and extracted with EtOAc (2×10 mL). Thecombined layer was washed with brine (10 mL), dried over Na₂SO₄,filtered and concentrated to give a residue, which was purified by flashcolumn (0-10% EtOAc in PE) to give N-8-22_1 (100 mg, 31%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.99-3.88 (m, 1H), 1.98-1.84 (m, 2H),1.69-1.57 (m, 6H), 1.52-1.45 (m, 2H), 1.44-1.28 (m, 3H), 1.26-1.17 (m,5H), 1.16-1.11 (m, 5H), 1.10-0.95 (m, 5H), 0.93-0.86 (m, 7H), 0.84-0.80(m, 4H), 0.69-0.62 (m, 4H).

2. DMP (224 mg, 0.53 mmol) was added to a solution of N-8-22_1 (100 mg,0.265 mmol) in DCM (10 mL). After stirring at 20° C. for 10 min, thereaction mixture was quenched with saturated NaHCO₃ solution (30 mL)until pH of the aqueous layer was about 9. The mixture was filtered. TheDCM layer was separated and the aqueous phase was extracted with DCM (20mL). The combined organic phase was washed with saturated Na₂S₂O₃ (3×40mL), sat.NaHCO₃ (40 mL), brine (40 mL), dried over Na₂SO₄, filtered,concentrated and purified by combi-flash (0-20% of EtOAc in DCM) to giveN-8-22_2 (80 mg, 80%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.54-2.42 (m, 1H), 2.09 (s, 3H), 1.94-1.87 (m,1H), 1.71-1.59 (m, 4H), 1.54-1.45 (m, 3H), 1.44-1.30 (m, 4H), 1.29-1.16(m, 6H), 1.15-1.07 (m, 5H), 1.06-0.92 (m, 4H), 0.91-0.79 (m, 7H),0.74-0.61 (m, 4H).

3. MeMgBr (0.353 mL, 1.06 mmol, 3M in ether) was added to a solution ofN-8-22_2 (80 mg, 0.213 mmol) in THF (5 mL) under N₂. After stirring at20° C. for 30 minutes, the reaction mixture was quenched with saturatedaqueous NH₄Cl (30 mL) solution and extracted with EtOAc (20 mL×3). Thecombined organic layer was washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated under vacuum to give a crude product, whichwas purified by silica gel column (0-10% of EtOAc in PE) to afford 26 (7mg, 8%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.08-2.01 (m, 1H), 1.97-1.86 (m, 1H),1.69-1.57 (m, 6H), 1.53-1.45 (m, 3H), 1.40-1.27 (m, 5H), 1.26-1.17 (m,8H), 1.14 (s, 3H), 1.13-1.01 (m, 3H), 0.99-0.92 (m, 5H), 0.91-0.85 (m,4H), 0.82 (s, 3H), 0.70 (s, 3H), 0.67-0.60 (m, 1H).

LCMS Rt=1.240 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₆ H₄₃ [M+H-2H₂O]⁺ 355, found 355.

Example 27: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-methylpentan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-H-cyclopenta[a]phenanthren-3-ol(27)

1. To a solution of N-014-012_4 (300 mg, 0.8366 mmol) in THF (20 mL) wasadded a solution of isopropylmagnesium chloride (1.25 mL, 2.50 mmol, 2M) drop-wise at 0° C. over a period of 30 mins under N₂, during whichthe temperature was maintained below 0° C. The reaction mixture wasstirred at 20° C. for another 2 hours to give a suspension. The reactionmixture was added saturated aq. NH₄Cl (15 mL) and stirred for 20 min,then the mixture was extracted with EtOAc (3×10 mL). The combinedorganic phase was washed with brine (2×10 mL), dried over anhydrousNa₂SO₄, filtered and concentrated to obtain N-014-001_1 (360 mg, crude)as a solid, ¹H NMR showed the desired product, and was used directly forthe next step.

2. X1 (150 mg, 0.37 mmol) was purified by SFC (Column: Chiralpak AS-H250*30 5 u; Condition: 0.1% NH₃H₂O EtOH; Begin B: 20%; End B: 20%;FlowRate (ml/min): 65) to obtain 37 (Peak 2, 46 mg, 31%) and 27 (Peak 1,27 mg, 18%) as a solid.

37:

27:

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.26 (m, 1H), 3.42-3.34 (m, 1H),2.43-2.34 (m, 1H), 2.06-1.91 (m, 3H), 1.90-1.75 (m, 2H), 1.74-1.66 (m,2H), 1.63-1.58 (m, 3H), 1.54-1.26 (m, 11H), 1.22-1.04 (m, 3H), 1.03-0.99(m, 3H), 0.97-0.93 (m, 7H), 0.92-0.87 (m, 3H), 0.86-0.77 (m, 3H), 0.70(s, 3H).

LCMS Rt=1.228 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₇H₄₅O [M+H-H₂O]⁺ 385, found 385.

SFC Rt=2.440 min in 10 min chromatography, OJ_3_EtOH_DEA_5_40_25ML(“Column: Chiralcel OJ-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B:ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40%for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Columntemp.: 35° C.″), 97.38% de.

Example 28: Synthesis of(3S,5S,8R,9R,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-13-methylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(28)

1. Pd/C (dry, 200 mg) was added to a solution of 44 (200 mg, 0.480 mmol)in MeOH/THF (10 mL/10 mL) under Ar. The suspension was degassed undervacuum and purged with H₂ three times. The mixture was stirred under H₂(50 psi) at 50° C. for 48 hrs to give a black suspension. The reactionmixture was filtered through a pad of Celite and washed with THF (100mL). The filtrate was concentrated to give 28 (30 mg, 15%) as a solidand 82 (30 mg, 15%) as a solid.

28:

¹H NMR (400 MHz, CDCl3) δ 3.63-3.61 (m, 1H), 1.98-1.76 (m, 4H),1.72-1.55 (m, 7H), 1.55-1.47 (m, 4H), 1.46-1.23 (m, 6H), 1.22-0.97 (m,11H), 0.92-0.78 (m, 12H), 0.76-0.54 (m, 5H).

LCMS Rt=1.298 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESIcalcd. For C₂₈H₄₇ [M+H-2H₂O]+ 383, found 383.

Example 29: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(29)

1. A solution of 1-bromo-3-methylbutane (11.7 g, 78 mmol) in THF (8 mL)was added dropwise to a suspension of Mg (4.35 g, 179 mmol) and I₂ (20mg) in THF (2 mL) at 60° C. The mixture was stirred at 60° C. for 1 hr.The mixture was diluted with THF (10 mL) and used directly. Freshlyprepared isopentylmagnesium bromide (19.5 mL, 3.9 M in THF, 76 mmol) wasadded to a solution of S-200-INT_5E (1.0 g, 2.78 mmol) in THF (5 mL)under N₂ at 0° C. The mixture was stirred at 0° C. for 1 hr. NH₄Cl (20mL, sat. aq.) was added to the mixture. The mixture was extracted withEtOAc (2×30 mL). The combined organic phase was washed with brine (100mL), dried over Na₂SO₄, concentrated under vacuum, purified by silicagel (PE/EtOAc=20/1 to 10/1), and re-crystallized from CH₃CN (10 mL) togive 77 (255 mg, 21%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.26 (m, 1H), 3.66-3.59 (m, 1H),2.42-2.32 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.58 (m, 4H), 1.55-1.38 (m,10H), 1.38-1.19 (m, 5H), 1.19-1.00 (m, 8H), 1.00-0.81 (m, 13H), 0.69 (s,3H).

LCMS Rt=1.306 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. for C₂₉H₄₉O [M+H-H₂O]⁺ 413, found 413.

2. Pd(OH)₂ (dry, 20%, 50.0 mg) was added to a solution of 77 (100 mg,232 umol) in THF (10 mL) and MeOH (10 mL) under Ar. The suspension wasdegassed under vacuum and purged with H₂ three times. The mixture wasstirred under H₂ (50 psi) at 50° C. for 16 hrs. The reaction mixture wasfiltered through a pad of Celite and washed with THF (3×10 mL). Thefiltrate was concentrated. The residue was purified by silica gelchromatography (PE/EtOAc=20/1) to afford 29 (7.00 mg, 7%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.68-3.60 (m, 1H), 1.96-1.88 (m, 2H),1.68-1.60 (m, 3H), 1.53-1.47 (m, 7H), 1.39-1.23 (m, 13H), 1.16-0.95 (m,7H), 0.90-0.86 (m, 12H), 0.83 (s, 3H), 0.66-0.63 (m, 4H).

LCMS Rt=1.603 min in 2.0 min chromatography, 30-90 AB_ELSD, purity 97%,MS ESI calcd. for C₂₉H₄₉ [M+H-H₂O]⁺ 397, found 397.

Example 30: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(30)

1. Benzoic acid (2.03 g, 16.7 mmol) and triphenylphosphine (6.57 g, 25.1mmol) were added to a solution of S-500-2-10 (3.5 g, 8.39 mmol) in THF(30 mL) at 25° C. under N₂. The mixture was stirred at 25° C. for 20mins. DIAD (5.07 g, 25.1 mmol) was added at 0° C. under N₂. The mixturewas stirred at 0° C. for 20 mins then warmed to 25° C. and stirred for 1h. The reaction mixture was quenched with water (100 mL) and extractedwith MTBE (3×30 mL). The combined organic phase was washed with brine(60 mL), dried over Na₂SO₄, filtered, concentrated in vacuum to give acrude, which was purified by flash column (0-10% EtOAc in PE) to give300 mg crude product S-500-2-9_1 as an oil, which was used directly forthe next step.

2._NaOH (1.14 g in 3 mL H₂O, 28.7 mmol) was added to a solution ofS-500-2-9_1 (300 mg, 0.576 mmol) in THF (5 mL) and MeOH (3 mL). Themixture was stirred at 50° C. for 16 hrs. The mixture was quenched withsat. NH₄Cl (20 mL) and extracted with EtOAc (3×10 mL). The combinedorganic phase was washed with brine (30 mL), dried over Na₂SO₄,filtered, concentrated and purified by combi-flash (0-10% of EtOAc inPE) to give 30 (12 mg, 5%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.28 (m, 1H), 3.63-3.59 (m, 1H),2.44-2.40 (m, 1H), 2.05-1.90 (m, 3H), 1.80-1.62 (m, 4H), 1.61-1.58 (m,3H), 1.56-1.30 (m, 9H), 1.28-1.03 (m, 10H), 1.01 (s, 3H), 0.99-0.85 (m,10H), 0.69 (s, 3H).

LCMS t_(R)=1.260 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₈H₄₅ [M+H-2H₂O]⁺ 381, found 381.

Example 31: Synthesis of(1R,3R,4S)-4-((3S,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1-phenylpentane-1,3-diol(31)

The synthesis of 31 can be found in Example 13.

31:

¹H NMR (400 MHz, CDCl₃) δ 7.45-7.28 (m, 5H), 5.02-4.81 (m, 1H),4.18-3.98 (m, 1H), 3.35 (brs, 1H), 2.47 (brs, 1H), 2.15-1.72 (m, 8H),1.53-1.31 (m, 8H), 1.30-1.03 (m, 8H), 0.99-0.89 (m, 4H), 0.89-0.78 (m,4H), 0.75-0.60 (m, 4H).

LCMS Rt=1.327 min in 2 min chromatography, 10-80AB_2MIN_E, purity 100%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.929 min in 10 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.4% de.

Example 32: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-phenylbutan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(32)

1. t-BuOK (4.64 g, 41.4 mmol) was added to a suspension of Me₃SI (8.44g, 41.4 mmol) in anhydrous THF (50 mL) at 20° C. under nitrogen. Themixture was stirred at 20° C. for 1 hr, and N-005_5 (5 g, 13.8 mmol) wasadded. The resulting mixture was warmed to 45° C. and stirred for 4 hrs.The reaction mixture was cooled to room temperature, quenched with water(50 mL), extracted with EtOAc (2×50 mL). The combined organic layer wasdried over anhydrous sodium sulfate, filtered and concentrated. Theresidue was purified by column chromatography on silica gel (0˜10% EtOAcin PE) to give N-005_001 (2.7 g, 52%) as a solid.

LCMS Rt=1.324 min in 2.0 min chromatography, 30-90AB_E, purity 92%, MSESI calcd. for C₂₅H₄₁O [M+H-H₂O]⁺ 357, found 357.

2. CuI (10.1 mg, 0.0534 mmol) and PhMgBr (1 M in THF, 2.66 mL, 2.66mmol) were added to a solution of N-005_001 (200 mg, 0.534 mmol) inanhydrous THF (20 mL) was at 0° C. under nitrogen. The mixture waswarmed to 15° C. gradually and stirred for 16 hrs. The reaction mixturewas quenched with aqueous NH₄Cl (20 mL), extracted with EtOAc (2×10 mL).The combined organic layer was dried over anhydrous sodium sulfate,filtered and concentrated. The residue was purified by columnchromatography on silica gel (0˜5% of EtOAc in PE) to give NA-6-5/6 (190mg, 79%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 7.35-7.27 (m, 2H), 7.25-7.18 (m, 3H),3.95-3.81 (m, 1H), 2.87-2.39 (m, 2H), 2.04-1.92 (m, 1H), 1.89-1.80 (m,1H), 1.71-1.58 (m, 4H), 1.56-1.43 (m, 6H), 1.41-1.27 (m, 5H), 1.26-1.18(m, 4H), 1.18-1.08 (m, 2H), 1.06-0.96 (m, 5H), 0.92-0.79 (m, 8H),0.73-0.55 (m, 4H).

3. N-6-5/6 (190 mg, 0.420 mmol) was separated by prep. HPLC (Column:YMC-Actus Triart C18 100*30 mm*5 um; condition: water (0.05% HCl)-ACN;Gradient: 90-100% B; Flow rate: 25 mL/min) to give 93 (56 mg, 30%) as asolid and 32 (12 mg, 6%) as a solid.

32:

¹H NMR (400 MHz, CDCl₃) δ 7.36-7.29 (m, 2H), 7.25-7.19 (m, 3H),3.89-3.83 (m, 1H), 2.79-2.72 (m, 1H), 2.49-2.40 (m, 1H), 2.05-1.98 (m,1H), 1.92-1.79 (m, 2H), 1.72-1.51 (m, 9H), 1.44-1.31 (m, 5H), 1.30-1.09(m, 7H), 1.08-0.96 (m, 5H), 0.92-0.81 (m, 7H), 0.74-0.63 (m, 4H).

LCMS Rt=1.343 min in 2.0 min chromatography, 30-90AB, purity 100%, MSESI calcd. for C₃₁H₄₇O [M+H-H₂O]⁺ 435, found 435.

Example 33: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-(3-methyloxetan-3-yl)butan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(33)

1. To a solution of N-014-005_1 (10 g, 97.9 mmol) in DCM (100 mL) wasadded 1-methyl-1H-imidazole (16.0 g, 195 mmol) and TEA (19.7 g, 195mmol) at 25° C. TsCl (37.1 g, 195 mmol) was added into the solution. Thereaction mixture was stirred at 25° C. for 2 hours. The mixture waswashed with water (2×100 mL), brine (100 mL), dried over Na2SO4,filtered and concentrated under vacuum to give N-014-005_2 (25 g, crude)as a light yellow solid, which was purified by column chromatography onsilica gel (0˜15% of EtOAc in PE) to give N-014-005_2 (23.6 g, 95%) as awhite solid.

¹H NMR (400 MHz, CDCl₃) δ 7.80-7.68 (m, 2H), 7.41-7.26 (m, 2H),3.40-3.29 (m, 4H), 4.12-4.00 (s, 2H), 2.44 (s, 3H), 1.28 (s, 3H).

2. To a solution of N-014-005_2 (10 g, 39.0 mmol) in acetone (100 mL)was added LiBr (13.5 g, 156 mmol). The mixture was stirred at 65° C. for1 hrs. The mixture was quenched with water (200 mL) at 0° C. andextracted with hexane (3×200 mL). The combined organic phase was washedwith brine (50 mL), dried over Na2SO4, filtered and concentrated to giveN-014-005_3 (2.54 g, crude) as yellow liquid.

¹H NMR (400 MHz, CDCl₃) δ 4.50-4.30 (m, 4H), 3.64 (s, 2H), 1.58 (s, 1H),1.43 (s, 3H).

3. To a suspension of Mg (807 mg, 33.2 mmol) and I₂ (1 mg) in THF (2 mL)was added solution of N-014-005_3 (2.5 g, 15.1 mmol) in THF (8 mL) dropwise under N₂ at 50˜55° C. The mixture was stirred at 55° C. for 1 h.The mixture was diluted with THF (10 mL) and used in the next stepdirectly without monitored. To a solution of N-14-12_4 (1.01 g, 2.83mmol) in THF (10 mL) was added fresh prepared 3-[(bromomagnesio)methyl]-3-methyloxetane (15 mmol in 20 mL of THF) at 0° C. The mixturewas stirred at 15° C. for 4 h. To the mixture was added NH₄Cl (20 mL,10% aq.). The mixture was extracted with EtOAc (30 mL). The organiclayer was separated and concentrated in vacuum. The residue was purifiedby flash column (0˜30% of EtOAc in PE) to give a mixture (190 mg, 15%)as a white solid, which was purified by SFC (Column: AD (250 mm*30 mm, 5um), Condition: 0.1% NH₃H₂O ETOH, Gradient: from 50% to 50%, FlowRate(ml/min): 60 mL/min, 25° C.) to afford 33 (Peak 1, 110 mg, 9%) and 70(Peak 2, 30 mg, impure) as a white solid. The impure 70 (30 mg, impure)was purified by column chromatography on silica gel (15% of EtOAc in PE)to give 70 (10 mg, 5%) as a white solid.

33:

¹H NMR (400 MHz, CDCl₃) δ 5.30-5.26 (m, 1H), 4.59-4.70 (m, 1H),4.50-4.48 (m, 1H), 4.36-4.33 (m, 1H), 3.83 (s, 1H), 2.40-2.33 (m, 1H),2.10-1.50 (m, 17H), 1.49-1.35 (m, 9H), 1.30-0.80 (m, 13H), 0.68 (s, 3H).

LCMS Rt=1.069 min in 3 min chromatography, 30-90AB_2MIN_E.M, purity100%, MS ESI calcd. for C₂₉H₄₉O₃ [M+H]⁺ 445, found 445.

Example 34: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-methylpentan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(34)

1. N-8-7_1 (500 mg, 1.38 mmol) in THF (5 mL) was added to a solution ofisopropylmagnesium chloride (708 mg, 3.44 mL, 2M in THF) in THF (5 mL)at 0° C. The mixture was stirred at 25° C. for 4 hrs. To the mixture wasadded NH₄Cl (20 mL, 10% aq.). The mixture was extracted with EtOAc (2×30mL). The organic layer was separated and concentrated in vacuum to givea residue. The residue was purified by silica gel chromatography elutedwith PE/EtOAc=3/1 to afford N-8-11_1 (170 mg, 30%) as a solid. Theimpure product (120 mg) was further purified by silica gelchromatography eluted with PE/EtOAc=3/1 to afford N-8-11_1 (50 mg, 42%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.20-3.10 (m, 1H), 2.00-1.80 (m, 2H),1.70-1.60 (m, 2H), 1.55-1.40 (m, 4H), 1.39-0.95 (m, 19H), 0.90-0.80 (m,15H), 0.70-0.60 (m, 5H).

2. DMP (457 mg, 1.08 mmol) was added to a solution of N-8-11_1 (220 mg,0.543 mmol) in DCM (5 mL). After stirring at 25° C. for 10 min, thereaction mixture was quenched with saturated NaHCO₃ aqueous (50 mL)until pH of the aqueous layer became about 9. The mixture was filtered.The DCM layer was separated and the aqueous phase was extracted with DCM(100 mL). The combined organic phase was washed with saturated Na₂S₂O₃aqueous (3×100 mL), sat. NaHCO₃ (100 mL), brine (40 mL), dried overNa₂SO₄, filtered and concentrated to give crude N-8-11_2 (140 mg, 64%)as a solid.

LCMS Rt=1.300 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity100%, MS ESI calcd. for C₂₇H₄₅O [M+H-H₂O]⁺ 385, found 385.

3. NaBH₄ (1.17 g, 17.3 mmol) was added five times, every five minutes,to a solution of N-8-11_2 (140 mg, 0.347 mmol) in MeOH (2 mL) and THF (2mL). The mixture was stirred at 15° C. for 30 minutes. The mixture wasquenched with sat.NH₄Cl (50 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was dried over Na₂SO₄, filtered, concentrated andpurified by combi-flash (25% of EtOAc in PE) to give N-8-11_1 (140 mg,impure). N-8-11_1 was purified by combi-flash (25% of EtOAc in PE) togive 34 (50 mg, impure) as a solid and 65 (10 mg, impure) as a solid.

4. N-8-11_1 (50 mg, 0.123 mmol, impure) was purified by combi-flash (25%of EtOAc in PE) to give 34 (30 mg, impure) as a solid.

N-8-11_1 (30 mg, 0.0741 mmol, impure) was purified by combi-flash (25%of EtOAc in PE) to give 34 (9 mg, 30%) as a solid.

34:

¹H NMR (400 MHz, CDCl₃) δ 3.18-3.07 (m, 1H), 1.98-1.81 (m, 2H),1.71-1.58 (m, 6H), 1.53-1.31 (m, 7H), 1.30-0.98 (m, 14H), 0.97-0.78 (m,14H), 0.70-0.60 (m, 4H).

LCMS Rt=4.387 min in 7.0 min chromatography, 30-90AB_7MIN_E, purity97.6%, MS ESI calcd for C₂₇H₄₅ [M+H-2H₂O]⁺ 369, found 369.

Example 35: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3-(methoxymethyl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(35)

1. NaOH (71.9 mg, 180 mmol) was added to a solution of N-4-4B (20 mg,0.0361 mmol) in THF/MeOH (2 mL) at 25° C. The reaction mixture warmed to50° C. and stirred for 1 h. The reaction mixture was cooled and water(20 mg) was added. The aqueous phase was extracted with EtOAc (3×10 mL).The combined organic phase was washed with saturated brine (2×20 mL),dried over anhydrous Na₂SO₄. filtered and concentrated. The residue waspurified by flash column (0˜30% of EtOAc in PE) to provide 35 (8 mg,50%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 3.63-3.61 (m, 1H), 3.41-3.38 (m, 5H); 2.51 (s,1H); 1.97-1.81 (m, 1H), 1.71-1.54 (m, 8H), 1.51-1.48 (m, 4H), 1.25-1.10(m, 15H), 0.99-0.80 (m, 9H), 0.78-0.75 (m, 4H), 0.71-0.59 (m, 4H).

LCMS Rt=1.301 min in 2.0 min chromatography, 30-90 AB, purity 96%, MSESI calcd. for C₂₉H₄₈O [M+H-2H₂O]⁺ 413, found 413.

Example 36: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(36)

1. Pd(OH)₂ (200 mg) was added to a solution of 30 (100 mg, 0.239 mmol)in MeOH (10 mL). The mixture was stirred at 50° C. under H₂ (50 Psi).The mixture was filtered, concentrated and purified by combi-flash(0-10% of EtOAc in PE) to give 47 (21 mg, 21%) and 36 (1 mg, 1%) as asolid.

37:

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.55 (m, 1H), 2.05-1.77 (m, 3H),1.72-1.63 (m, 3H), 1.55-1.48 (m, 3H), 1.47-1.31 (m, 9H), 1.29-1.12 (m,13H), 1.11-1.00 (m, 3H), 0.96 (s, 3H), 0.93-0.87 (m, 9H), 0.67 (s, 3H).

LCMS t_(R)=1.296 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₈H₄₇ [M+H-2H₂O]⁺ 383, found 383.

Example 37: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-methylpentan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-H-cyclopenta[a]phenanthren-3-ol(37)

1. N-014-001_1 (150 mg, 0.37 mmol) was purified by SFC (Column:Chiralpak AS-H 250*30 5 u; Condition: 0.1% NH₃H₂O EtOH; Begin B: 20%;End B: 20%; FlowRate (ml/min): 65) to obtain 37 (Peak 2, 46 mg, 31%) and27 (Peak 1, 27 mg, 18%) as a solid. 014-001A:

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.28 (m, 1H), 3.18-3.09 (m, 1H),2.39-2.35 (m, 1H), 2.06-1.81 (m, 4H), 1.73-1.57 (m, 6H), 1.54-1.41 (m,8H), 1.40-1.26 (m, 3H), 1.24-1.11 (m, 3H), 1.10-0.97 (m, 6H), 0.96-0.92(m, 1H), 0.90-0.85 (m, 5H), 0.84-0.76 (m, 4H), 0.69 (s, 3H).

LCMS Rt=1.207 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₇H₄₅O [M+H-H₂O]⁺ 385, found 385.

SFC Rt=2.635 min in 10 min chromatography, OJ_3_EtOH_DEA_5_40_25ML(“Column: Chiralcel OJ-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B:ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40%for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Columntemp.: 35° C.″), 98.66% de.

Example 38: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxyhexan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(38)

1. Propylmagnesium bromide (3.34 mL, 6.69 mmol, 2M in THF) was slowlyadded to a solution of S-500-6-1_1 (800 mg, 2.23 mmol) in THF (30 mL) at0° C. After addition, the mixture was stirred at 15° C. for 1 hr. Themixture was quenched with sat. NH₄Cl (40 mL) and extracted with EtOAc(3×20 mL). The combined organic phase was washed with brine (2×30 mL),dried over Na₂SO₄, filtered and concentrated and purified by combi-flash(0-15% of EtOAc in PE) to give 72 (500 mg, 56%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.72-3.64 (m, 1H),2.41-2.31 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.69 (m, 1H), 1.62-1.54 (m,3H), 1.52-1.38 (m, 9H), 1.37-1.16 (m, 6H), 1.15-1.01 (m, 7H), 0.99-0.88(m, 7H), 0.87-0.82 (m, 3H), 0.68 (s, 3H).

LCMS Rt=4.979 min in 7.0 min chromatography, 30-90AB_E, purity 98.8%, MSESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

2. Pd(OH)₂ (300 mg, dry) was added to a solution of 72 (150 mg, 0.372mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 23 (9 mg, 6%) and 38 (43 mg,29%) as a solid.

38:

¹H NMR (400 MHz, CDCl₃) δ 3.71-3.62 (m, 1H), 1.99-1.82 (m, 2H),1.70-1.56 (m, 6H), 1.54-1.45 (m, 3H), 1.44-1.38 (m, 3H), 1.37-1.17 (m,10H), 1.16-1.01 (m, 5H), 1.00-0.85 (m, 11H), 0.82 (s, 3H), 0.70-0.60 (m,4H).

LCMS Rt=1.397 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₇H₄₅ [M+H-2H₂O]⁺ 369, found 369.

Example 39: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-(methoxymethyl)-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(39)

1. t-BuOH (600 mL) was charged into a three-neck round bottom flaskunder N₂ at 35° C., followed by adding t-BuOK (101 g, 905 mmol). Afterstirring at 35° C. for 30 mins, N-004-029_1 (50 g, 151 mmol) was addedto the above mixture and stirred at 35° C. for 1 hr. The reactionmixture was poured into 5% aqueous acetic acid (2 L), during which thetemperature was maintained below 10° C. Ice-water (1 L) was added. ThepH of the mixture was adjusted to about 7-8 with NaHCO₃ and filtered.The filter cake was dissolved in DCM (1.5 L). The combined organic phasewas washed with water (2×500 ml), brine (2×500 ml), dried over Na₂SO₄,filtered and concentrated in vacuum at below 35° C. to give N-004-029_2(45 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.32 (m, 1H), 3.71-3.58 (m, 1H),3.38-3.25 (m, 2H), 2.90-2.78 (m, 1H), 2.55-2.20 (m, 2H), 2.13-1.92 (m,3H), 1.90-1.59 (m, 5H), 1.46-1.14 (m, 10H), 1.12-0.96 (m, 6H), 0.72 (s,3H).

2. n-BuLi (108 mL, 272 mmol, 2.5 M in h-hexane) was added dropwise to amixture of Me₃SI (73.8 g, 362 mmol) in anhydrous THF (300 mL) at 0° C.under N₂. After stirring at 0° C. for 30 mins, a solution of N-004-029_2(30 g, 90.7 mmol) in anhydrous THF (600 mL) was added at −40° C. Themixture was stirred −40° C. for 2 hours and at 25° C. for 16 hours. Thereaction mixture was poured into ice-water (1 L). The aqueous phase wasextracted with EtOAc (2×500 mL). The combined organic phase was washedwith brine (2×500 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated, the residue was purified by flash column (0˜20% of EtOAcin PE) to give N-004-029_3 (1.8 g, 6%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.25 (m, 1H), 3.66-3.61 (m, 1H),3.39-3.31 (m, 1H), 2.93-2.86 (m, 1H), 2.59-2.53 (m, 1H), 2.20-1.93 (m,4H), 1.89-1.14 (m, 15H), 1.12-0.90 (m, 9H), 0.71 (s, 3H).

3. MeONa (5.61 g, 104 mmol) was added to a solution of N-004-029_3 (1.8g, 5.22 mmol) in MeOH (20 mL) at 25° C. under N₂. After stirring at 50°C. for 12 hrs, water (100 mL) was added into the mixture and stirred for10 mins. The aqueous phase was extracted with EtOAc (2×80 mL). Thecombined organic phase was washed with saturated brine (2×50 mL), driedover anhydrous Na₂SO₄, filtered and concentrated. The residue waspurified by flash column (0˜30% of EtOAc in PE) to give N-004-029_4 (1.5g, 76%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.28 (m, 1H), 3.70-3.54 (m, 1H),3.40-3.35 (m, 6H), 3.28-3.16 (m, 2H), 2.40-2.35 (m, 1H), 2.09-1.90 (m,5H), 1.87-1.57 (m, 11H), 1.34-1.06 (m, 10H), 0.70 (s, 3H).

4. DMP (2.53 g, 5.97 mmol) was added to a solution of N-004-029_4 (1.5g, 3.98 mmol) in DCM (30 mL) at 25° C. and after stirring at 25° C. for30 min. The reaction mixture was quenched with saturated NaHCO₃ (50 mL)at 25° C. DCM (50 mL) was added into the mixture and stirred for 10 min.The DCM phase was separated and washed with saturated Na₂S₂O₃ aqueous(2×50 mL), brine (2×50 mL), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by flash column (5˜25% of EtOAcin PE) to give N-004-029_5 (0.6 g, 40%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.59-9.57 (m, 1H), 5.32-5.29 (m, 1H), 3.37 (s,3H), 3.30-3.15 (m, 2H), 2.44-2.31 (m, 2H), 2.13-1.40 (m, 16H), 1.27-1.02(m, 10H), 0.73 (s, 3H).

5. CsF (607 mg, 4.00 mmol) was added to a solution of N-004-029_5 (0.6g, 1.60 mmol) in anhydrous THF (20 mL) at 0° C. After stirring for 20min, TMSCF₃ (568 mg, 4.00 mmol) was added at 0° C. and the mixture wasstirred for 1 hr. TBAF.3H₂O (2.02 g, 6.40 mmol) was added into themixture, which was stirred at 50° C. for 1 hr. The reaction mixture waspoured into ice-water (50 mL). The aqueous phase was extracted withEtOAc (2×80 mL). The combined organic phase was washed with saturatedbrine (2×80 mL), dried over anhydrous Na₂SO₄, filtered and concentrated.The residue was purified by flash column (0˜30% of EtOAc in PE) to giveN-004-029A (450 mg, 63%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.29 (m, 1H), 4.11-3.99 (m, 1H), 3.37 (s,3H), 3.30-3.19 (m, 2H), 2.54 (s, 1H), 2.43-2.36 (m, 1H), 2.26-1.82 (m,7H), 1.78-1.61 (m, 5H), 1.34-0.80 (m, 15H), 0.75-0.67 (m, 3H).

6. The N-004-029A (0.45 g, 1.01 mmol) was purified by SFC (Column: AD(250 mm*30 mm, 5 um), Condition: 0.1% NH₃H₂O ETOH, Begin B: 30%, End B:30%) to afford 39 (PK1: 120 mg, 26.7%) as a white solid and 95 (PK2: 200mg, 44.5%) as a white solid.

The structure of 39 was confirmed by NOE.

39:

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.29 (m, 1H), 4.06-3.99 (m, 1H), 3.37 (s,3H), 3.30-3.19 (m, 2H), 2.54 (s, 1H), 2.43-2.36 (m, 1H), 2.25-2.19 (m,1H), 2.15-2.07 (m, 1H), 2.04-1.60 (m, 9H), 1.55-1.34 (m, 5H), 1.25-0.88(m, 11H), 0.70 (s, 3H).

LCMS Rt=1.078 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₄H₃₄F₃O [M-CH₅O₂]⁺ 395, found 395.

Example 40: Synthesis of(3S,5S,8R,9R,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-13-methylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(40)

1. Pd(OH)₂ (100 mg, dry) was added to a solution of 3 (30 mg, 0.072mmol) in MeOH/THF (5 mL/5 mL) at 25° C. under Ar. The reaction wasstirred at 50° C. under H₂ (50 Psi) for 48 h. The mixture was filteredand the filtrate was concentrated in vacuum to give crude product, whichwas purified by a silica gel column (PE/EtOAc=10/1-5/1) to give 41 (10mg, 33%) as a solid.

¹H NMR (400 MHz, CDCl3) δ 3.62-3.59 (m, 1H), 1.99-1.91 (m, 1H),1.88-1.76 (m, 2H), 1.74-1.61 (m, 6H), 1.46-1.29 (m, 6H), 1.26-1.09 (m,9H), 1.08-0.99 (m, 6H), 0.95-0.78 (m, 14H), 0.74-0.58 (m, 5H).

LCMS Rt=1.491 min in 2 min chromatography, 30-90 AB, purity 99%, MS ESIcalcd. For C₂₈H₄₇ [M+H-2H₂O]⁺ 383, found 383.

Example 41: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxyhept-5-yn-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(41)

1. n-BuLi (13.8 mL, 34.5 mmol, 2.5 M in hexane) was added dropwise to asolution of but-2-yne (1.86 g, 34.5 mmol) in THF (100 mL) at −20° C. Thesolution was stirred for 2.5 hrs −20° C. and then it was cooled to −78°C., N-8-7_1 (5.0 g, 13.8 mmol) in THF (100 mL) was added. The solutionwas stirred for 30 mins at this temperature, then at −20° C. for 1 hr,and followed by 20° C. for 18 hrs. The resulting gel was quenched bypouring into sat. NH₄Cl (100 mL), followed by extraction with EtOAc(2×150 mL). The combined organic layers were washed with water (40 mL)and brine (40 mL), dried over Na₂SO₄, filtered and purified by flashcolumn eluting with 0˜20% of EtOAc in PE to give N-8-7_2B (1 g, crude)as an oil and N-8-7_2B (1.7 g, crude) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 3.90-3.75 (m, 1H), 2.45-2.35 (m, 1H),2.25-2.05 (m, 3H), 1.95-1.85 (m, 2H), 1.85-1.40 (m, 8H), 1.40-1.20 (m,9H), 1.20-0.75 (m, 18H), 0.65 (s, 4H).

SFC Peak 1: Rt=3.008 min in 10 min chromatography,AD_3_EtOH_DEA_5_40_25ML (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 umMobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% ofB in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate:2.5 mL/min Column temp.: 35° C.).

2. The crude N-8-7_2B (250 mg, 0.868 mmol) was further purified by SFC(column: AD (250 mm*30 mm, 10 um)), gradient: 35-35% B (A=0.1% NH₃/H₂O,B=EtOH), flow rate: 60 mL/min) to give 41 (peak 2, 81 mg, 33%) as asolid and 68 (peak 1, 78 mg, 31%) as a solid.

41:

¹H NMR (400 MHz, CDCl₃) δ 3.82-3.70 (m, 1H), 2.79-2.08 (m, 2H),2.00-1.90 (m, 1H), 1.80 (s, 4H), 1.78-1.69 (m, 1H), 1.69-1.42 (m, 10H),1.40-1.31 (m, 4H), 1.31-1.18 (m, 4H), 1.18-0.92 (m, 6H), 0.92-0.85 (m,7H), 0.82 (s, 3H), 0.66 (s, 3H).

LCMS Rt=1.206 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₈H₄₅O [M+H-H₂O]⁺ 397 found 397.

SFC Rt=5.823 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

Example 42: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(42)

1. 4-methylbenzenesulfonic acid (2.70 g, 15.7 mmol) was added to asolution of N-4-1_1 (50 g, 157 mmol) in MeOH (500 mL) at 25° C. Themixture was stirred at 65° C. for 1 hr. The reaction mixture was cooledto 25° C. and TEA (2.16 mL, 15.7 mmol) was added. The mixture wasstirred for 0.5 h. The precipitate was collected by filtration andwashed with methanol (2×100 mL) to give N-4-1_2 (50 g, crude) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 3.25-3.05 (m, 6H), 2.60-2.40 (m, 1H),2.20-2.05 (m, 4H), 2.00-1.95 (m, 1H), 1.90-1.80 (m, 1H), 1.75-1.50 (m,6H), 1.49-1.05 (m, 12H), 1.04-0.95 (m, 1H), 0.78 (s, 3H), 0.59 (s, 3H).

2. t-BuOK (23.0 g, 205 mmol) was added to a solution ofbromo(methyl)triphenylphosphorane (73.2 g, 205 mmol) in THF (500 mL) at25° C. The mixture was heated to 45° C. and stirred for 1 hr. N-4-1_2(50 g, 137 mmol) was added. The mixture was stirred at 45° C. for 2 hrs.The mixture was quenched with NH₄Cl (200 mL) and extracted with THF(3×100 mL). The organic layer was washed brine (200 mL), dried overNa₂SO₄ and filtered to give a mixture (50 g, 500 mL), which was used innext step without further purification.

3. Aqueous HCl (207 mL, 1 M in water) was added to a solution of N-4-1_3(50 g, 138 mmol) in THF (500 mL). The mixture was stirred at 25° C. for0.5 h. The mixture was filtered and the filter cake was dissolved in DCM(200 mL) and washed with brine (100 mL), dried over anhydrous Na₂SO₄,filtered and concentrated in vacuum to afford N-4-1_4 (39 g, 90%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.45-2.20 (m, 3H),2.15-2.00 (m, 3H), 1.90-1.65 (m, 8H), 1.60-1.50 (m, 2H), 1.45-1.05 (m,8H), 1.00 (s, 3H) 0.90-0.85 (m, 1H), 0.80-0.75 (m, 1H), 0.58 (s, 3H).

4. CsF (25.9 g, 171 mmol) and TMSCF₃ (24.3 g, 171 mmol) were added to asolution of N-4-1_4 (27 g, 85.8 mmol) in THF (200 mL). The mixture wasstirred at 10° C. for 1 hr. Water (10 mL) and TBAF.3H₂O (30 g) wereadded to the mixture. The mixture was stirred at 30° C. for another 2hrs. The mixture was concentrated in vacuum. The residue was dissolvedin EtOAc (500 mL), washed with water (2×500 mL), dried over Na₂SO₄,filtered, concentrated in vacuum and purified by flash column (DCM/EtOAc(1:1) in PE, 0˜10%) to give N-4-1_5 (27 g, 82%) and N-4-1_5A (3.5 g,11%) as a solid.

N-4-1_5:

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.12-1.94 (m, 3H),1.89-1.78 (m, 2H), 1.75 (s, 3H), 1.72-1.60 (m, 5H), 1.58-1.48 (m, 2H),1.45-1.09 (m, 10H), 1.01-0.89 (m, 1H), 0.85 (s, 3H), 0.78-0.68 (m, 1H),0.56 (s, 3H).

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.09-1.99 (m, 1H),1.89-1.78 (m, 2H), 1.75 (s, 3H), 1.72-1.52 (m, 9H), 1.45-1.06 (m, 10H),1.00-1.81 (m, 2H), 0.79 (s, 3H), 0.56 (s, 3H).

5. 9-BBN dimer (29 g, 119 mmol) was added to a solution of N-4-1_5 (23g, 59.8 mmol) in THF (250 mL) and the mixture was stirred at 40° C.under N₂ for 16 hrs. Ethanol (34.3 mL, 598 mmol) and NaOH (119 mL, 5 M,598 mmol) were added to the reaction mixture. The mixture became clear.H₂O₂ (59.8 mL, 10 M, 598 mmol) was added dropwise at 25° C. and theinner temperature was raised to reflux (70° C.). The mixture was cooledto 30° C. after the addition. To the mixture was added Na₂SO₃ (100 mL,20% aq.). The organic layer was separated and poured into water (800mL). A solid formed. The mixture was filtered and the solid was washedwith water, dried under vacuum and triturated with MeCN (250 mL) to givea solid. The solid was triturated from MeOH/water (250 mL/12.5 mL) at60° C. and filtered after cooled to 15° C. The solid was dried undervacuum to give N-4-1_6 (16.4 g, 68%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.69-3.60 (m, 1H), 3.39-3.29 (m, 1H),2.09-2.01 (m, 1H), 1.99-1.92 (m, 1H), 1.87-1.75 (m, 2H), 1.72-1.43 (m,7H), 1.42-1.07 (m, 11H), 1.03 (d, J=6.8 Hz, 3H), 1.01-0.86 (m, 3H), 0.85(s, 3H), 0.73-0.69 (m, 1H), 0.67 (s, 3H).

6. Water (223 mg, 12.4 mmol) and DMP (10.5 g, 24.8 mmol) were added to asuspension of N-4-1_6 (5 g, 12.4 mmol) in DCM (200 mL). The mixture wasstirred at 15° C. for 15 mins. The mixture was washed withNaHCO₃/Na₂S₂O₃ (200 mL/200 mL, sat.) twice, dried over Na₂SO₄, filteredand concentrated in vacuum to give N-4-1_7 (4.5 g, 90%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.60-9.51 (m, 1H), 2.40-2.30 (m, 1H),2.12-1.78 (m, 5H), 1.75-1.59 (m, 4H), 1.57-1.15 (m, 11H), 1.14-0.84 (m,8H), 0.78-0.63 (m, 5H).

7. A solution of 1-bromo-3-methylbutane (2.79 g, 18.5 mmol) in THF (8mL) was added dropwise to a suspension of Mg (899 mg, 37 mmol) and 12 (1mg) in THF (2 mL) under N₂ at 50-55° C. The mixture was stirred at 55°C. for 1 hr to give the isopentylmagnesium bromide solution. The freshlyprepared isopentylmagnesium bromide (18.5 mmol in 10 mL of THF) wasadded to a solution of N-4-1_7 (0.5 g, 1.24 mmol) in THF (5 mL) at 0° C.The mixture was stirred at 15° C. for 2 hrs. To the mixture was addedNH₄Cl (20 mL, 10% aq.). The mixture was extracted with EtOAc (2×30 mL).The combined organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuum to give N-4-2 (0.6 g, crude) as a solid.

8. Water (1 drop) and DMP (1.06 g, 2.52 mmol) were added to a solutionof N-4-2 (0.6 g, 1.26 mmol) in DCM (20 mL) at 15° C. The mixture wasstirred at 15° C. for 1 h. The mixture was washed with NaHCO₃/Na₂S₂O₃(20 mL/20 mL, sat.) twice, dried over Na₂SO₄, filtered and concentratedin vacuum to give N-4-1_8 (0.6 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.59-2.30 (m, 3H), 2.11-1.78 (m, 4H),1.75-1.36 (m, 13H), 1.35-0.98 (m, 11H), 0.91-0.82 (m, 10H), 0.78-0.70(m, 1H), 0.67 (s, 3H).

9. NaBH₄ (0.96 g, 25.4 mmol) was added in portions to a solution ofN-4-1_8 (0.6 g, 1.27 mmol) in THF (10 mL) and MeOH (5 mL) at 15° C. Themixture was stirred at 15° C. for 30 mins. To the mixture was addedNH₄Cl (50 mL, 10%). The mixture was extracted with EtOAc (2×50 mL). Thecombined organic layer was dried over Na₂SO₄, filtered and concentratedunder vacuum and purified by flash column (0˜15% EtOAc in PE) to giveimpure 42 and 85. N42 was triturated from MeCN (10 mL) at 15° C. anddried in vacuum to give 42 (153 mg, 25%) as a solid. 85 was purified byflash column (0˜15% EtOAc in PE) to give an oil, which was treated withMeCN (5 mL) and water (5 mL), and concentrated under vacuum to give 85(70 mg, 12%) as a solid.

42:

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.55 (m, 1H), 2.01-1.78 (m, 6H),1.71-1.59 (m, 4H), 1.51-1.15 (m, 16H), 1.09-1.02 (m, 3H), 0.92-0.81 (m,13H), 0.72-0.61 (m, 4H).

LCMS Rt=1.378 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₈H₄₆F₃O [M+H-H₂O]⁺ 455, found 455.

HPLC Rt=5.38 min in 10.0 min chromatography, 50-100_AB_E, purity 99.58%.

Example 43: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(43)

1. To a solution of 62 (160 mg, 0.373 mmol) in MeOH (2 mL) and THF (1mL) was added Pd(OH)₂ (0.2 g, <1% water). The solution was hydrogenatedunder 50 psi of hydrogen at 50° C. for 16 hrs. Then the mixture wasfiltered through a pad of celite and the filtrate was concentrated invacuum. The residue was purified by flash column (PE/EtOAc=10/1 to 5/1)to give 43 (27 mg, 17%) and 16 (117 mg, 73%) as a white solid.

43:

¹H NMR (400 MHz, CDCl3) δ.4.05-3.99 (m, 1H), 1.99-1.81 (m, 5H),1.79-1.72 (m, 1H), 1.70-1.56 (m, 3H), 1.53-1.35 (m, 7H), 1.35-1.07 (m,12H), 1.04-1.02 (m, 3H), 0.97 (s, 3H), 0.92 (t, J=8 Hz, 3H), 0.70 (s,3H).

LCMS Rt=1.271 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₄₀F₃O [M+H-H₂O]⁺ 413, found 413.

Example 44: Synthesis of(3S,8R,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-13-methyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(44)

1. A solution of 1-bromo-3-methylbutane (4 g, 26.4 mmol) in THF (27 mL)was added dropwise to a suspension of Mg (947 mg, 39.5 mmol) and 12(33.5 mg, 0.132 mmol) in THF (3 mL) at 60° C. The mixture was stirred at60° C. for 1 hr. Freshly prepared isopentylmagnesium bromide (30 mL,0.88 M in THF, 26.4 mmol) was added to a solution of S-500-15-2_1 (800mg, 2.32 mmol) in THF (2 mL) under N₂ at 0° C. The mixture was stirredat 0° C. for 1 h. To the mixture was added NH4Cl (50 mL, sat. aq.), themixture was extracted with EtOAc (2×50 mL). The combined organic phasewas washed with brine (100 mL), dried over Na2SO4, filtered andconcentrated in vacuum to give crude product which was purified bysilica gel (PE/EtOAc=10/1 to 5/1) to give 44 (720 mg, 75%) as a whitesolid.

1H NMR (400 MHz, CDCl3) δ 5.40-5.38 (m, 1H), 3.63-3.61 (m, 1H),2.23-2.21 (m, 1H), 2.10-1.74 (m, 7H), 1.69-1.58 (m, 2H), 1.54-1.34 (m,8H), 1.33-1.00 (m, 11H), 0.95-0.75 (m, 14H), 0.70 (s, 3H).

LCMS Rt=1.289 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESIcalcd. For C28H₄₅ [M+H-2H2O]+ 381, found 381.

Example 45: Synthesis of(3S,5R,8R,9R,10S,13S,14S,17R)-3-(methoxymethyl)-13-methyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(45)

1. To a solution of G-020-004_1 (100 g, 364 mmol) in anhydrous methanol(1 L) was added TsOH (6.26 g, 36.4 mmol). The mixture was stirred at 60°C. for 18 hrs. The reaction mixture was concentrated to remove most ofthe solvent, neutralized with Et₃N (3.7 g), diluted with EtOAc (600 mL),washed with water (500 mL) and brine (500 mL). The organic layer wasconcentrated to give G-020-004_2 (133 g, crude) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 3.20 (s, 3H), 3.14 (s, 3H), 2.63-2.39 (m, 2H),2.14-2.03 (m, 2H), 1.97-1.89 (m, 2H), 1.86-1.77 (m, 3H), 1.64-1.60 (m,2H), 1.56-1.49 (m, 3H), 1.47-1.42 (m, 2H), 1.40-1.32 (m, 2H), 1.29-1.23(m, 3H), 1.16-1.06 (m, 2H), 0.87 (s, 3H).

2. To a suspension of Ph₃PEtBr (308 g, 830 mmol) in anhydrous THF (700mL) under nitrogen at 20° C. was added t-BuOK (93.1 g, 830 mmol). Afterstirring at 20° C. for 1 hr, a solution of G-020-004_2 (133 g, 415 mmol)in anhydrous THF (300 mL) was added to the mixture. The resultingmixture was warmed to 50° C. and stirred for 4 hrs. The reaction mixturewas cooled, quenched with water (400 mL) and sat. NH₄Cl (300 mL),stirred for 30 min. The organic layer was separated, and the water phasewas extracted with THF (300 mL). The combined organic layer was useddirectly in next step.

3. To a solution of G-020-004_3 (137 g, 412 mmol, theoretical) in THF(1.3 L) was added aqueous HCl (1 M, 618 mL, 618 mmol). After stirring at20° C. for 1 hr, the reaction mixture was quenched with saturated NaHCO₃(800 mL) and extracted with EtOAc (2×500 mL). The combined organic layerwas dried over anhydrous sodium sulfate, filtered and concentrated togive a solid (300 g). The solid was triturated from petroleum ether (800mL) for 18 hrs. The solid was filtered off, and the filter cake waswashed with petroleum ether (400 mL). The filtrate was concentrated togive a residue (117 g). The residue was purified by columnchromatography on silica gel (0˜10% of EtOAc in PE) to give N-004-023_5(70 g) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.17-5.09 (m, 1H), 2.65-2.55 (m, 1H),2.43-2.34 (m, 1H), 2.33-2.15 (m, 6H), 2.11-2.05 (m, 1H), 1.83-1.70 (m,2H), 1.68-1.64 (m, 4H), 1.63-1.59 (m, 2H), 1.58-1.46 (m, 3H), 1.42-1.25(m, 3H), 1.25-1.14 (m, 3H), 0.91 (s, 3H).

4. A stirred solution of trimethylsulfoxonium iodide (30.5 g, 139 mmol)and t-BuOK (15.5 g, 139 mmol) in DMSO (200 mL) was heated at 60° C. for1.0 h under N₂; N-004-023_5 (20 g, 69.8 mmol) was added to the reactionmixture and stirred at 60° C. for 10 mins. The reaction was treated withwater (1000 mL). The aqueous phase was extracted with EtOAc (2×500 mL).The combined organic phase was washed with water (2×500 mL), brine (300mL), dried over anhydrous Na₂SO₄, filtered and concentrated under vacuumto afford N-004-023_6 (20.5 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.13-5.10 (m, 1H), 2.62-2.60 (m, 2H),2.25-2.20 (m, 5H), 2.00-1.48 (m, 12H), 1.46-1.00 (m, 8H), 0.98-0.89 (m,4H).

5. MeONa (18.4 g, 341 mmol) was added to a solution of N-004-023_6 (20.5g, 68.2 mmol) in MeOH (500 mL) at 25° C. under N₂, The mixture wasstirred at 70° C. reflux for 16 h under N₂, The reaction was treatedwith water (500 mL). The aqueous phase was extracted with DCM (2×300mL). The combined organic phase was washed with saturated brine (2×300mL), dried over anhydrous Na₂SO₄, filtered and concentrate was purifiedby silica gel chromatography (PE/EtOAc=10/1 to 6/1) to affordN-004-023_7 (20 g, 88%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.12-5.06 (m, 1H), 3.38 (s, 3H), 3.19 (s, 2H),2.25-2.22 (m, 1H), 2.20-2.09 (m, 3H), 1.66-1.63 (m, 3H), 1.60-1.24 (m,14H), 1.22-1.00 (m, 6H), 0.87 (s, 3H).

6. 9-BBN dimer (29.2 g, 120 mmol) was added to a solution of N-004-023_7(20 g, 60.1 mmol) in THF (100 mL) at 0° C. under N₂. The solution wasstirred at 65° C. for 2 hrs. After cooling to 0° C., EtOH (34.9 mL, 601mmol) was added. Then a solution of NaOH (120 mL, 5M, 601 mmol) wasadded very slowly. After addition, H₂O₂ (68.0 g, 601 mmol, 30% in water)was added slowly and the inner temperature was maintained below 10° C.The mixture was stirred at 75° C. under N₂ for 1 hr. The mixture wasre-cooled to 25° C. The mixture was added to H₂O (2 L). The mixture wasstirred 30 mins. The precipitate was collected by filtration and washedwith H₂O (2×500 mL) to give N-004-023_8 (17.8 g, 85%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.70-3.55 (m, 1H), 3.38 (s, 3H), 3.19 (m, 2H),2.11-1.86 (m, 4H), 1.80-1.25 (m, 13H), 1.23-0.88 (m, 12H), 0.68 (s, 3H).

7. Silica gel (24 g) and PCC (24.5 g, 114 mmol) were added to asuspension of N-004-023_8 (20 g, 57.0 mmol) in DCM (500 mL) at 25° C.The mixture was stirred at 25° C. for 2 hrs. The mixture was filteredand the filtered cake was washed with DCM (2×100 mL). The combinedfiltrate was concentrated under vacuum. The residue was purified byflash column (0˜30% of EtOAc in PE) to give NA-004-023_9 (19 g, 95%) asa solid.

¹H NMR (400 MHz, CDCl₃) δ 3.39 (s, 3H), 3.20 (s, 2H), 2.60-2.52 (m, 1H),2.20-2.10 (m, 5H), 1.99-1.80 (m, 3H), 1.75-1.40 (m, 12H), 1.30-1.04 (m,7H), 0.61 (s, 3H).

8. t-BuOK (12.2 g, 54.5 mmol) was added to a suspension of MePPh₃Br(38.9 g, 109 mmol) in THF (300 mL) at 25° C. After addition, thereaction mixture was heated to 45° C. and stirred for 1 hr. ThenN-004-023_9 (19 g, 35.9 mmol) was added and the reaction mixture wasstirred at 45° C. for 16 hrs. The mixture was treated with NH₄Cl (100mL, sat. aq.). The organic layer was separated. The aqueous phase wasextracted with EtOAc (2×300 mL). The combined organic phase was washedwith saturated brine (2×200 mL), dried over anhydrous Na₂SO₄, filteredand concentrated. The residue was triturated from MeOH/H₂O (100 mL/100mL) at 25° C. to give N-004-023_10 (17 g, 90%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.68 (s, 1H), 3.39 (s, 3H), 3.19(s, 2H), 2.10-2.04 (m, 1H), 2.03-1.90 (m, 3H), 1.75-1.56 (m, 12H),1.49-1.25 (m, 4H), 1.22-0.89 (m, 8H), 0.57 (s, 3H).

9. 9-BBN dimer (29.5 g, 122 mmol) was added to a solution ofN-004-023_10 (17 g, 49.0 mmol) in anhydrous THF (300 mL) and stirred at0° C. for 30 min under N₂. The reaction mixture was warmed to 25° C.(room temperature) and stirred for 2 hrs. The reaction mixture wascooled. The mixture was quenched by EtOH (100 mL) at 0° C. NaOH (98.0mL, 490 mol, 5M in water) was added very slowly. After addition, H₂O₂(44.5 mL, 490 mmol, 11M) was added slowly until the inner temperature nolonger rises and during which the temperature was maintained below 30°C. The mixture was stirred at 50° C. for another 1 hr. then the mixturewas cooled, treated with water (2 L) and stirred for 30 min. Thesuspension was filtration in vacuum to give N-004-023_11 (17 g, crude)as a solid. The N-004-023_11 (17 g, 46.6 mmol) was triturated fromMeOH/H₂O (100/100 mL) at 25° C. and stirred for 1 hr. The suspension wasfiltered under vacuum to obtain N-004-023_11 (14 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.79-3.66 (m, 1H), 3.50-3.37 (m, 4H), 3.28 (s,2H), 2.43 (s, 1H), 2.26-1.98 (m, 3H), 2.18-2.12 (m, 1H), 1.95-1.60 (m,11H), 1.34-1.04 (m, 14H), 0.76 (s, 3H).

10. DMP (9.24 g, 21.8 mmol) was added to a solution of N-004-023_11 (4g, 10.9 mmol) in DCM (80 ml) at 25° C. One drop of water was added tothe mixture and was stirred for 30 mins. The reaction mixture wasquenched with saturated NaHCO₃, aqueous pH=7-8 at below 10° C. The DCMphase in the filtrate was separated and washed with saturatedNaHCO₃/Na₂S₂O₃ aqueous (1:1, 2×50 mL). The organic phase was washed withsaturated brine (2×50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuum to give N-004-023_12 (1.8 g, 46%) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 9.57-9.55 (m, 1H), 3.38 (s, 3H), 3.20 (s, 2H),2.39-2.26 (m, 1H), 2.17-2.06 (m, 1H), 2.05-1.75 (m, 4H), 1.74-1.53 (m,8H), 1.85-1.00 (m, 15H), 0.74 (s, 3H).

11. CsF (1.86 g, 12.3 mmol) was added to a solution of N-004-023_12 (1.8g, 4.96 mmol) in anhydrous THF (20 mL) at 0° C. After stirring at 0° C.for 20 min, TMSCF₃ (1.74 g, 12.3 mmol) was added at 0° C. and stirredfor 1 hr, then TBAF.3H₂O (6.25 g, 19.8 mmol) was added. The mixturereaction was warmed to 50° C. and stirred for another 1 hr. The reactionmixture was poured into ice-water (50 mL) and stirred for 10 min. Theaqueous phase was extracted with EtOAc (2×80 mL). The combined organicphase was washed with saturated brine (2×80 mL), dried over anhydrousNa₂SO₄, filtered and concentrated. The residue was purified by flashcolumn (0˜20% of EtOAc in PE) to give N-004-023_13 (1.2 g, 56%) as anoil.

¹H NMR (400 MHz, CDCl₃) δ 4.03-3.98 (m, 1H), 3.39 (s, 3H), 3.20 (s, 2H),2.17-1.80 (m, 7H), 1.73-1.41 (m, 10H), 1.28-0.95 (m, 13H), 0.71 (s, 3H).

12. DMP (2.34 g, 5.54 mmol) was added to a solution of N-004-023_13 (1.2g, 2.77 mmol) in DCM (30 ml) at 25° C. After stirring at 25° C. for 30mins, the reaction mixture was quenched with saturated NaHCO₃ (30 mL),aqueous pH=7-8 at below 10° C. Then DCM (30 mL) was added and themixture was stirred for 10 min. The suspension was filtered. The DCMphase in the filtrate was separated and washed with saturatedNaHCO₃/Na₂S₂O₃ aqueous (1:1, 2×50 mL). The organic phase was washed withsaturated brine (2×50 mL). dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuum to give N-004-023_13A (1.2 g, crude) as an oil.

¹H NMR (400 MHz, CDCl₃) δ 3.39 (s, 3H), 3.20 (s, 2H), 3.05-2.95 (m, 1H),1.91-1.51 (m, 10H), 1.46-1.20 (m, 10H), 1.17-0.96 (m, 8H), 0.72 (s, 3H).

13. NaBH₄ (210 mg, 5.56 mmol) was added to a solution of N-004-023_13A(1.2 g, 2.78 mmol) in MeOH (5 mL) at 0° C. and stirred for 30 min. Aftertreating with MeOH/H₂O (20/20 mL), the mixture was stirred for 10 min.The aqueous phase was extracted whit EtOAc (2×50 mL). The combineorganic phase was washed with saturated brine (2×50 mL), drive overanhydrous Na₂SO₄, filtered and concentrated. The residue was purified byflash column (0˜20% of EtOAc in PE) to give 48 (57 mg, 5%) as a solidand 45 (200 mg, impure) as a solid. The 45 (200 mg, 0.462 mmol) waspurified by flash column (0˜30% of EtOAc in PE) to give 45 (120 mg, 10%)as a solid.

45:

¹H NMR (400 MHz, CDCl₃) δ 4.01-3.98 (m, 1H), 3.38 (s, 3H), 3.19 (s, 2H),2.15-2.10 (m, 1H), 2.05-1.80 (m, 5H), 1.72-1.55 (m, 5H), 1.54-1.34 (m,6H), 1.31-1.20 (m, 4H), 1.16-0.95 (m, 9H), 0.71 (s, 3H).

LCMS Rt=1.129 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₄H₃₈F₃ [M−HO₃]⁺ 383, found 383.

Example 46: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3R)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(46)

1. A solution of (bromomethyl)cyclopentane (2.25 g, 13.8 mmol) in THF (8mL) was added dropwise to a suspension of Mg (662 mg, 27.6 mmol) and I₂(70 mg, 0.276 mmol) in THF (3 mL) at 75° C. The mixture was stirred at75° C. for 1 hr. After cooling, a solution of S-500-6-1_1 (1 g, 2.78mmol) in THF (30 mL) was added slowly at 15° C. After addition, themixture was stirred at 15° C. for 2 hrs, quenched with sat.NH₄Cl (40 mL)and sat. citric acid (20 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was washed with brine (2×30 mL), dried overNa₂SO₄, filtered and concentrated and purified by combi-flash (0-15% ofEtOAc in PE) to give a mixture of S-500-6-13_1 and an isomer at the 22position (900 mg, 73%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.23 (m, 1H), 3.77-3.67 (m, 1H),2.40-2.30 (m, 1H), 2.07-1.89 (m, 4H), 1.88-1.69 (m, 4H), 1.67-1.59 (m,4H), 1.55-1.26 (m, 15H), 1.16-1.05 (m, 5H), 1.05-1.00 (m, 4H), 0.99-0.81(m, 8H), 0.68 (s, 3H).

2. DMP (1.72 g, 4.06 mmol) was added to a solution of S-500-6-13_1 (900mg, 2.03 mmol) in DCM (30 mL). After that, the reaction mixture wasstirred at 15° C. for 10 min. The reaction mixture was quenched withsaturated NaHCO₃ aqueous (50 mL) until pH of the aqueous layer becameabout 9. The mixture was filtered. The DCM layer was separated and theaqueous phase was extracted with DCM (20 mL). The combined organic phasewas washed with saturated aqueous Na₂S₂O₃ (3×40 mL) and sat. NaHCO₃ (40mL), brine (40 mL), dried over Na₂SO₄, filtered and concentrated to givecrude S-500-6-13_2 (900 mg, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.30-5.25 (m, 1H), 2.54-2.42 (m, 2H),2.41-2.33 (m, 1H), 2.30-2.17 (m, 1H), 2.06-1.90 (m, 3H), 1.87-1.78 (m,2H), 1.73-1.66 (m, 2H), 1.65-1.35 (m, 15H), 1.33-1.21 (m, 2H), 1.17-0.92(m, 13H), 0.88-0.82 (m, 3H), 0.69 (s, 3H).

3. NaBH₄ (3.46 g, 102 mmol) was added five times, every five minutes, toa solution of S-500-6-13_2 (900 mg, 2.04 mmol) in MeOH (5 mL) and THF (5mL). The mixture was stirred at 15° C. for 30 minutes, quenched withsat. NH₄Cl (50 mL) and extracted with EtOAc (3×20 mL). The combinedorganic phase was dried over Na₂SO₄, filtered, concentrated and purifiedby combi-flash (0-15% of EtOAc in PE) to give impure 46 (120 mg) as asolid, which was separated by SFC ((column: AD (250 mm*30 mm, 5 um),gradient: 45-45% B (A=0.05% NH₃/H₂O, B=MeOH), flow rate: 60 mL/min) togive pure 46 (100 mg, 84%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.76-3.67 (m, 1H),2.40-2.33 (m, 1H), 2.08-1.91 (m, 4H), 1.90-1.78 (m, 2H), 1.77-1.55 (m,10H), 1.54-1.31 (m, 9H), 1.26-1.22 (m, 2H), 1.22-1.05 (m, 6H), 1.03 (s,3H), 1.01-0.89 (m, 5H), 0.89-0.82 (m, 3H), 0.69 (s, 3H).

LCMS Rt=1.474 min in 2.0 min chromatography, 30-90AB_E, purity 99%, MSESI calcd. for C30H₄₉O [M+H-H₂O]⁺ 425, found 425.

Example 47: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(47)

1. Pd(OH)₂ (200 mg) was added to a solution of 30 (100 mg, 0.239 mmol)in MeOH (10 mL). The mixture was stirred at 50° C. under H₂ (50 Psi).The mixture was filtered, concentrated and purified by combi-flash(0-10% of EtOAc in PE) to give 47 (21 mg, 21%) and 36 (1 mg, 1%) as awhite solid.

47:

¹H NMR (400 MHz, CDCl₃) δ 3.68-3.54 (m, 1H), 2.02-1.90 (m, 1H),1.76-1.57 (m, 6H), 1.54-1.27 (m, 10H), 1.26-1.21 (m, 7H), 1.20-1.08 (m,5H), 1.07-0.95 (m, 3H), 0.94-0.83 (m, 10H), 0.81 (s, 3H), 0.72-0.60 (m,4H).

LCMS t_(R)=1.290 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₈H₄₇ [M+H-2H₂O]⁺ 383, found 383.

Example 48: Synthesis of(3S,5R,8R,9R,10S,13S,14S,17R)-3-(methoxymethyl)-13-methyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(48)

NaBH₄ (210 mg, 5.56 mmol) was added to a solution of N-004-023_13A (1.2g, 2.78 mmol) in 1. MeOH (5 mL) at 0° C. and stirred for 30 min. Aftertreating with MeOH/H₂O (20/20 mL), the mixture was stirred for 10 min.The aqueous phase was extracted whit EtOAc (2×50 mL). The combineorganic phase was washed with saturated brine (2×50 mL), drive overanhydrous Na₂SO₄, filtered and concentrated. The residue was purified byflash column (0˜20% of EtOAc in PE) to give 48 (57 mg, 5%) as a solidand 45 (200 mg, impure) as a solid. The 45 (200 mg, 0.462 mmol) waspurified by flash column (0˜30% of EtOAc in PE) to give 45 (120 mg, 10%)as a solid.

48:

¹H NMR (400 MHz, CDCl₃) δ 4.07-4.01 (m, 1H), 3.39 (s, 3H), 3.19 (s, 2H),2.30-2.22 (m, 1H), 2.14-2.05 (m, 1H), 2.00-1.80 (m, 4H), 1.72-1.57 (m,6H), 1.49-1.20 (m, 9H), 1.18-0.95 (m, 9H), 0.68 (s, 3H).

LCMS Rt=1.085 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₄H₃₈F₃ [M−HO₃]⁺ 383, found 383.

Example 49: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3S)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(49)

The synthesis of 49 is described in Example 4.

49:

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.85-3.77 (m, 1H),2.40-2.32 (m, 1H), 2.07-1.87 (m, 4H), 1.76-1.69 (m, 1H), 1.66-1.55 (m,5H), 1.53-1.42 (m, 7H), 1.41-1.31 (m, 5H), 1.30-1.12 (m, 8H), 1.11-1.05(m, 3H), 1.03 (s, 3H), 1.01-0.92 (m, 2H), 0.91-0.82 (m, 12H), 0.68 (s,3H).

LCMS Rt=1.718 min in 2.0 min chromatography, 30-90AB_E, purity 98%, MSESI calcd. for C₃₃H₅₃ [M+H-2H₂O]₊ 449, found 449.

Example 50: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxybutan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(50)

1. MeMgBr (0.83 mL, 2.49 mmol, 3M in ether) was added dropwise to asolution of N-8-7_1 (300 mg, 0.832 mmol) in THF (20 mL) at 0° C. underN₂. After stirring at 20° C. for 30 minutes, the reaction was quenchedwith sat. NH₄Cl (50 mL) and extracted with EtOAc (2×10 mL). The combinedphase was washed with brine (10 mL), dried over Na₂SO₄, filtered,concentrated and purified by flash column (step 2) (0-10% EtOAc in PE)to give 50 (40 mg, 29%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.98-3.89 (m, 1H), 1.99-1.84 (m, 2H),1.69-1.56 (m, 6H), 1.54-1.45 (m, 2H), 1.43-1.29 (m, 6H), 1.28-1.17 (m,4H), 1.17-1.12 (m, 4H), 1.12-0.94 (m, 5H), 0.92-0.84 (m, 7H), 0.82 (s,3H), 0.68-0.61 (m, 4H).

LCMS Rt=3.428 min in 7.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₅ H₄₁ [M+H-2H₂O]⁺ 341, found 341.

Example 51: Synthesis of3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-(methoxymethyl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(51)

1. S-500-6-29_2 (999 mg, 1.22 M in THF, 4.87 mmol) was added dropwise toa solution of N-8-1_1 (210 mg, 05576 mmol) in THF (2 mL) at 25° C. underN₂. After stirring at 25° C. for 16 hrs, the reaction mixture wasquenched with saturated NH₄Cl (10 mL) and extracted with ethyl acetate(3×10 mL). The combined organic layer was washed with brine (30 mL),dried over Na₂SO₄, filtered and concentrated in vacuum to give a crudeproduct, which was purified by flash column (0˜15% of EtOAc in PE) for 2times to give impure product (30 mg). The impure product was furtherpurified by ELSD prep-HPLC (column: Phenomenex Synergi C18 150*30 mm*4um), gradient: 90-95% B (A=water (0.05% HCl), B=MeCN), flow rate: 25mL/min) to give pure 51 (4 mg, 1.4% yield) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.84-3.76 (m, 1H), 3.45-3.32 (m, 5H),2.62-2.39 (m, 1H), 1.99-1.85 (m, 2H), 1.73-1.62 (m, 4H), 1.53-1.40 (m,7H), 1.39-1.31 (m, 5H), 1.30-1.21 (m, 7H), 1.20-1.13 (m, 4H), 1.12-1.10(m, 5H), 0.99-0.93 (m, 1H), 0.89-0.86 (m, 6H), 0.85 (s, 3H), 0.83 (s,3H), 0.68-0.61 (m, 4H).

LCMS Rt=5.669 min in 7.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₅O [M+H-2H₂O]⁺ 467, found 467.

Example 52: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-H-cyclopenta[a]phenanthren-3-ol(52)

1. NaOH solution (974 mg in 6 mL H₂O, 16.8 mmol) was added to a solutionof S-500-2-15_1 (900 mg, 1.68 mmol) in THF (10 mL) and MeOH (5 mL). Themixture was heated at 50° C. for 16 hrs. The reaction mixture wasquenched with sat. NH₄Cl (60 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was washed with brine (60 mL), dried over Na₂SO₄,filtered, concentrated, and purified by combi-flash (0-15% of EtOAc inPE) to give 210 mg of a solid, which was purified by SFC (column: AD(250 mm*30 mm, 5 um), gradient: 35-35% B (A=0.1% NH₃/H₂O, B=MeOH), flowrate: 80 mL/min) to give 52 (150 mg, 68%) as a solid.

¹H NMR (400 MHz, CDCl3) δ 5.30-5.26 (m, 1H), 3.64-3.58 (m, 1H),2.40-2.30 (m, 1H), 2.02-1.92 (m, 3H), 1.80-1.58 (m, 7H), 1.56-1.31 (m,9H), 1.30-1.05 (m, 8H), 1.03 (s, 3H), 1.02-0.96 (m, 2H), 0.95-0.86 (m,9H), 0.85-0.80 (m, 3H), 0.69 (s, 3H).

LCMS t_(R)=1.335 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₉H₄₇ [M+H-2H₂O]⁺ 395, found 395.

Example 53: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-3-(trifluormethyl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(53)

1. To a solution of N-004-027_1 (1.5 g, 3.76 mmol) in anhydrous THF (40mL) was added CsF (1.42 g, 9.40 mmol) at 0° C. After stirring at 0° C.for 20 min, TMSCF₃ (1.33 g, 9.40 mmol) was added at 0° C. and stirredfor 30 min. The color becomes light yellow. TBAF.3H₂O (4.74 g, 15.0mmol) was added and stirred at 50° C. for 30 min. The reaction mixturewas poured into ice-water (100 mL). The aqueous phase was extracted withEtOAc (2×100 mL). The combined organic phase was washed with saturatedbrine (2×100 mL), dried over anhydrous Na₂SO₄, filtered and concentratedto give a mixture of isomers (1.45 g, crude) as a yellow solid, whichwas purified by flash column (0˜15% of EtOAc in PE) to give 53 (340 mg,24%) as a white solid and 1 (200 mg, 14%) as a white solid.

53:

¹H NMR (400 MHz, CDCl₃) δ 5.38-5.36 (m, 1H), 4.06-3.94 (m, 1H), 2.49 (s,2H), 2.09-1.58 (m, 13H), 1.48-0.85 (m, 14H), 0.73 (s, 3H).

LCMS Rt=1.134 min in 2 min chromatography, 30-90AB_2MIN_E, purity 99%,

MS 50-100_1_4 min·m, for C₂₄H₃₃F₆O [M+H-H₂O]⁺ 451, found 451.

Example 54: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6,6-dimethylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(54)

1. NaBH₄ (1.77 g, 46.8 mmol) was added in portions to a solution ofS-500-6-1_3 (520 mg, 1.17 mmol) in THF (5 mL) and MeOH (10 mL) at 15° C.The mixture was stirred at 15° C. for 20 min. The mixture was quenchedwith NH₄Cl (20 mL, sat. aq.) and extracted with EtOAc (50 mL). Theorganic layer was separated and concentrated under vacuum to give amixture which was separated by flash column (0˜15% EtOAc in PE) to giveS-500-6-1 (300 mg, impure) and 54 (170 mg, impure).

The impure 54 (220 mg, impure) was purified by flash column (0˜15% EtOAcin PE) to give a solid. The solid was dissolved in MeCN (50 mL) at 60°C. and concentrated under vacuum to give 54 (120 mg, 23%) as a solid.

54:

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.24 (m, 1H), 3.62-3.52 (m, 1H),2.42-2.31 (m, 1H), 2.11-1.90 (m, 3H), 1.72-1.35 (m, 15H), 1.29-1.08 (m,8H), 1.03 (s, 3H), 1.01-0.96 (m, 2H), 0.93 (d, J=6.8 Hz, 3H), 0.90 (s,9H), 0.85 (t, J=7.6 Hz, 3H), 0.70 (s, 3H).

LCMS Rt=5.463 min in 7.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₃₀H₄₉ [M+H-2H₂O]⁺ 409, found 409.

Example 55: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(55)

1. A solution N-8-7_1 (300 mg, 0.832 mmol) in THF (5 mL) was added to asolution of PhMgBr (1.38 mL, 3 M in ether, 4.15 mmol) in THF (10 mL) at0° C., then the reaction mixture was stirred at 0° C. for 3 hours. Next,the reaction mixture was stirred at 25° C. for 5 hours. The reactionmixture was quenched by water (10 mL) at 0° C. The solution was filteredand the filter cake was washed with EtOAc (10 mL). The aqueous phase wasextracted with EtOAc (3×15 mL). The combined organic phase was washedwith saturated brine (2×10 mL), dried over anhydrous Na₂SO₄, filteredand concentrated. The residue was purified by silica gel chromatography(PE/EtOAc=20/1 to 1/1) to afford 59 and 19 (200 mg, crude) as a solid.The crude product was purified by SFC (column: AD (250 mm*30 mm, 5 um)),gradient: 25-25% B (A=0.1% NH₃/H2O, B=EtOH), flow rate: 60 mL/min) togive 55 (Peak2, 55 mg, 15%) and 19 (Peak1, 21 mg, 6%) as solids.

55:

¹H NMR (400 MHz, CDCl₃) δ 7.38-7.28 (m, 4H), 7.25-7.20 (m, 1H),64.95-4.90 (m, 1H), 2.13-2.01 (m, 1H), 1.98-1.88 (m, 1H), 1.77-1.59 (m,6H), 1.57-1.43 (m, 6H), 1.43-0.93 (m, 13H), 0.91-0.85 (m, 3H), 0.83 (s,3H), 0.76-0.72 (m, 3H), 0.68 (s, 4H).

LCMS Rt=1.239 min in 2.0 min chromatography, 30-90AB_2 min., purity100%, MS ESI calcd. For C₃₀H₄₃ [M-2H₂O+H]⁺ 403, found 403.

SFC Rt=1.192 min in 3 min chromatography, OJ_3_EtOH_DEA_5_40_25ML, 99%de.

Example 56: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(56)

1. To a solution of 81 (1 g, 3.26 mmol) in MeOH (30 mL) and THF (10 mL)was added Pd(OH)₂ (1 g, <1% water). Then the mixture was hydrogenatedunder 50 psi at 50° C. for 48 hrs. The mixture was filtered through apad of celite without monitor and the filtrate was concentrated invacuum. The residue was purified by flash column (PE/EtOAc=10/1 to 5/1)to give 56 (331 mg, 33%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 4.09-3.99 (m, 1H), 2.18-2.13 (m, 1H),1.99-1.78 (m, 4H), 1.75-1.59 (m, 3H), 1.50-1.3 (m, 7H), 1.34-1.22 (m,6H), 1.21-1.00 (m, 10H), 0.96 (s, 3H), 0.94-0.89 (m, 3H), 0.67 (s, 3H).

LCMS Rt=1.184 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₅H₄₀F₃O [M+H-H₂O]⁺ 413, found 413.

Example 57: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3,10,13-trimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(57)

1. A suspension of LiCl (13.9 g, 329 mmol, anhydrous) in THF (500 mL,anhydrous) was stirred at 10° C. for 30 mins under N₂. FeCl₃ (27.8 g,172 mmol, anhydrous) was added at 10° C. The mixture was cooled to −30°C. To the mixture was added MeMgBr (209 mL, 3M in diethyl ether) droppedat −30° C. After stirring at −30° C. for 10 mins, S-500-2-12_1 (50 g,157 mmol) was added at −30° C. The mixture was stirred at −15° C. for 2hours and quenched with citric acid (500 mL, 10% aq.). The mixture wasextracted with EtOAc (3×800 mL). The combined organic phase was washedwith saturated brine (300 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under vacuum to give crude product, which was purified by asilica gel column (PE/DCM/EtOAc=1/1/1) to give S-500-2-12_2 (50 g, 86%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.57-2.48 (m, 1H), 2.23-2.13 (m, 1H),2.06-1.78 (m, 3H), 1.64-1.25 (m, 14H), 1.24-1.01 (m, 10H), 0.96 (s, 3H),0.74 (s, 1H), 0.60 (s, 3H).

2. To a suspension of PPh₃MeBr (79.7 g, 244 mmol) in THF (400 mL) wasadded t-BuOK (25.1 g, 224 mmol) at 20° C. After stirring at 40° C. for30 min, a solution of S-500-2-12_2 (50 g, 150 mmol) in THF (100 mL) wasadded at 40° C. and the reaction mixture was stirred at 40° C. for 1 hr.The reaction mixture was poured into 50 g of ice and stirred for 15minutes. The organic layer was separated and the water phase wasextracted with EtOAc (3×50 mL). The combined organic phase concentratedin vacuum to give an oil. The crude product was triturated in MeOH/H₂O(200 mL/200 mL) and filtered to give S-500-2-12_3 (55 g, 88%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.69 (s, 1H), 2.06-1.79 (m, 4H),1.75 (s, 3H), 1.73-1.58 (m, 4H), 1.56-1.25 (m, 9H), 1.22 (s, 3H),1.21-1.02 (m, 6H), 1.01-0.94 (s, 3H), 0.93-0.74 (m, 1H), 0.55 (s, 3H).

3. The solution of S-500-2-12_3 (55 g, 166 mmol) in THF (500 mL) wasadded 9-BBN dimer (60.7 g, 249 mmol) was stirred at 25° C. under N₂ for1 hr, a solid was formed. To the reaction mixture was added ethanol(95.3 mL, 1.66 mol) and NaOH (166 mL, 5 M, 830 mmol). The mixture turnedclear. H₂O₂ (132 mL, 10 M, 1.32 mol) was added dropwise at 25° C. andthe inner temperature was raised to reflux (75° C.). The mixture wascooled after addition and stirred for 16 hrs, a solid was formed. To themixture was added Na₂S₂O₃ (500 mL, 20% aq.) and water (1 L) at 25° C.The mixture was stirred for 1 hr. After the stirrer was turned off, aclear lower layer and a upper suspension layer were formed. The clearlower layer was discarded. To the upper suspension layer was added water(2 L). The mixture was stirred for 15 mins. The mixture was filtered togive S-500-2-12_4 (50 g, impure) as a solid. S-500-2-12_4 (50 g, 143mmol, impure) was triturated in EtOH/H₂O (90 mL/10 mL) at 100° C. for 2hours, then cooled to 15° C. and filtered to give S-500-2-12_4 (38 g,impure) as a solid. S-500-2-12_4 (38 g, 109 mmol, impure) was trituratedin EtOH/H₂O (45 mL/5 mL) at 100° C. for 2 hours, then cooled to 15° C.and filtered to give S-500-2-12_4 (28 g, 43%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.67-3.59 (m, 1H), 3.39-3.32 (m, 1H),2.01-1.75 (m, 4H), 1.69-1.58 (m, 3H), 1.54-1.24 (m, 10H), 1.23-1.14 (m,9H), 1.09-1.02 (m, 5H), 0.96 (s, 3H), 0.74 (m, 1H), 0.67 (s, 3H).

4. To a solution of N-004-016_1 (10.0 g, 28.6 mmol) in DCM (100 mL) wasadded DMP (24.2 g, 57.2 mmol). Then H₂O (0.2 mL) was added to themixture. After that, the reaction was stirred at 25° C. for 1 hour. Tothe reaction mixture was added saturated aqueous NaHCO₃ (100 mL)solution. The mixture was filtered and the filter cake was washed withDCM (2×100 mL). The mixture was liquid was separated, and the waterphase was extracted with DCM (2×100 mL). The combined organic layer waswashed with saturated aqueous NaHCO₃/Na₂S₂O₃ (100 mL/100 mL) and brine(100 mL), dried over Na₂SO₄, filtered and concentrated in vacuum to givea white solid. The residue was purified by silica gel chromatography(PE/EtOAc=0 to 20%) to afford N-004-016_2 (3.5 g, 35%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 9.58-9.54 (m, 1H), 2.39-2.32 (m, 1H),1.96-1.77 (m, 4H), 1.69-1.31 (m, 14H), 1.23-1.16 (m, 6H), 1.14-1.02 (m,5H), 0.96 (s, 3H), 0.76-0.59 (m, 4H).

5. To a solution of N-004-016_2 (1.5 g, 4.32 mmol), CsF (328 mg, 2.16mmol) in THF (10 mL) was added TMSCF₃ (1.53 g, 10.8 mmol) at 0° C. Themixture was stirred at 25° C. for 1 hrs. To the mixture was addedTBAE₃H₂O (3.4 g, 10.8 mmol). The mixture was stirred at 25° C. for 2hrs. The mixture was quenched with water (20 mL) and extracted withEtOAc (2×30 mL). The combined organic phase was washed with brine (50mL), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum.The residue was purified by flash column (0˜15% of EtOAc in PE) toafford N-004-017_1 (700 mg, 39%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 4.07-3.96 (m, 1H), 2.21-2.09 (m, 1H),1.99-1.77 (m, 5H), 1.72-1.29 (m, 15H), 1.24-1.20 (m, 4H), 1.13-1.01 (m,5H), 0.96 (s, 3H), 0.89-0.84 (m, 1H), 0.76-0.64 (m, 3H), 0.60 (s, 1H).

6. To a solution of N-004-017_1 (700 mg, 1.68 mmol) in pyridine (5 mL)was added benzoyl chloride (354 mg, 2.52 mmol) and DMAP (102 mg, 0.84mmol) at 25° C. The mixture was heated to 60° C. and stirred for 10hours. The reaction mixture was diluted with EtOAc (10 mL), thenquenched with water (10 mL). The aqueous was extracted with EtOAc (2×20mL). The combined organic layer was washed with brine (20 mL), driedover Na₂SO₄, filtered and concentrated in vacuum to give a white solid.The residue was purified by flash column (0˜15% of EtOAc in PE) to giveN-004-017_2 (600 mg, 68%) as a white solid.

LCMS Rt=1.464 min in 2 min chromatography, 30-90AB_2MIN_E.M, purity 92%.

SFC condition: Peak 1: Rt=2.434 min and Peak 2: Rt=2.541 min in 8 minchromatography, OD_MEOH(DEA)_5_40_2,8ML_8MIN.M (Column: Chiralcel OD-3100×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: methanol (0.05% DEA)Gradient: from 5% to 40% of B in 4.5 min and hold 40% for 2.5 min, then5% of B for 1 min Flow rate: 2.8 mL/min Column temperature: 40° C.).

7. N-004-017_2 (600 mg, 1.15 mmol) was purified by SFC (column:Chiralcel OD (250 mm*30 mm, 5 um), gradient: 20-20% B (A=0.1% NH₃/H₂O,B=MeOH), flow rate: 60 mL/min) to give N-004-017_3 (Peak 2, 190 mg,impure, 31%), N-004-018_1 (Peak 1, 180 mg, 30%) as a white solid. Theimpure N-004-017_3 (190 mg, 0.36 mmol) was purified by SFC (column: OD(250 mm*30 mm, 5 um), gradient: 20-20% B (A=0.1% NH₃/H₂O, B=MeOH), flowrate: 60 mL/min) to give N-004-017_3 (100 mg, 53%) as a white solid.

N-004-018_1:

¹H NMR (400 MHz, CDCl3) δ 8.12-8.06 (m, 2H), 7.65-7.59 (m, 1H),7.53-7.46 (m, 2H), 5.68-5.58 (m, 1H), 2.15-2.03 (m, 2H), 1.97-1.69 (m,3H), 1.67-1.57 (m, 3H), 1.44-1.24 (m, 9H), 1.23-1.13 (m, 11H), 1.12-1.97(m, 3H), 0.94 (s, 3H), 0.72 (s, 3H).

LCMS Rt=1.525 min in 2 min chromatography, 30-90AB_2 min_220&254.1 cm,purity 100%.

SFC_D1 Rt=2.450 min in 8 min chromatography,OD_MEOH(DEA)_5_40_2,8ML_8MIN.M, 100% de.

N-004-017_3:

¹H NMR (400 MHz, CDCl3) δ 8.14-8.06 (m, 2H), 7.64-7.57 (m, 1H),7.52-7.44 (m, 2H), 5.63-5.52 (m, 1H), 2.11 (s, 1H), 2.06-1.77 (m, 6H),1.72-1.62 (m, 3H), 1.44-1.32 (m, 8H), 1.28-1.18 (m, 12H), 0.99-0.93 (m,4H), 0.65 (s, 3H).

LCMS Rt=1.529 min in 2 min chromatography, 30-90AB_2 min_220&254.1 cm,purity 100%.

SFC Rt=2.544 min in 8 min chromatography,OD_MEOH(DEA)_5_40_2,8ML_8MIN.M, 98% de.

8. To a solution of N-004-018_1 (180 mg, 0.34 mmol) in THF (3 mL) andMeOH (1.5 mL) and water (1.5 mL) was added KOH (96.5 mg, 1.72 mmol). Themixture was stirred at 60° C. for 16 hrs. The mixture was poured intowater (20 mL) and extracted with EtOAc (2×40 mL). The combined organiclayer was washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by flash column (0˜15% of EtOAcin PE) to give 57 (114 mg, 79%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 4.00 (brs, 1H), 2.03-1.77 (m, 8H), 1.69-1.61(m, 2H), 1.54-1.49 (m, 1H), 1.47-1.24 (m, 10H), 1.22 (s, 3H), 1.21-1.10(m, 4H), 1.08-1.01 (m, 4H), 0.96 (s, 3H), 0.69 (s, 3H).

LCMS Rt=1.169 min in 2 min chromatography, 30-90AB_2MIN_E.M, purity 95%,MS ESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

HPLC Rt=5.44 min in 10 min Ultimate C18 3*50 mm 3 um,30-90_AB_1.2ML_E.MET, purity 100%.

Example 58: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((1R,2S)-1-cyclopentyl-1-hydroxypropan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(58)

1. N-8-7_1 (500 mg, 1.38 mmol) in THF (5 mL) was added tocyclopentylmagnesium bromide (1.38 mL, 3 M in THF) at 0° C. under N₂.After stirring at 15° C. for 18 hrs, the reaction mixture was quenchedwith sat. NH₄Cl (10 mL), and extracted with EtOAc (2×10 mL). Thecombined organic layer was washed with brine (10 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by flash column(0-10% of EtOAc in PE) to give N-8-15_1 (170 mg, 29%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.39-3.22 (m, 1H), 2.00-1.81 (m, 4H),1.70-1.41 (m, 12H), 1.41-1.13 (m, 13H), 1.13-0.95 (m, 6H), 0.95-0.79 (m,11H), 0.65 (s, 3H).

2. DMP (0.881 g, 2.08 mmol) was added to a solution of N-8-15_1 (300 mg,0.696 mmol) in DCM (20 mL). After stirring at 15° C. for 10 min, thereaction mixture was quenched with sat.NaHCO₃ (10 mL). The mixture wasextracted with DCM (3×20 mL). The combined organic phase was washed withsaturated Na₂S₂O₃ (3×20 mL) and brine (20 mL), dried over Na₂SO₄,filtered, concentrated and purified by combi-flash (0-20% of EtOAc inPE) to give N-8-15_2 (240 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.98-2.89 (m, 1H), 2.62-2.55 (m, 1H),1.98-1.87 (m, 1H), 1.81-1.72 (m, 4H), 1.71-1.49 (m, 10H), 1.41-1.29 (m,4H), 1.29-1.19 (m, 6H), 1.14-0.98 (m, 9H), 0.94-0.87 (m, 4H), 0.82 (s,3H), 0.67 (m, 5H).

3. NaBH₄ (550 mg, 14.5 mmol) was added to a mixture of N-8-15_2 (240 mg,0.559 mmol) in MeOH (3 mL) and THF (2 mL). The mixture was stirred at15° C. for 0.5 h. Another batch of NaBH₄ (550 mg, 14.5 mmol) was added.The reaction mixture was stirred for another 1 h. To the reactionmixture was added water (5 mL). The resulting mixture was extracted withEtOAc (2×10 mL). The combined organic layer was washed with brine (10mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column (0˜5% EtOAc in PE) to give and 58 (7 mg, 5%) asa solid and 78 (50 mg, impure) was further purified by flash column(0˜5% of EtOAc in PE) to give 78 (17 mg, 12%) as a solid.

58:

¹H NMR (400 MHz, CDCl₃) δ 3.64-3.59 (m, 1H), 2.09-1.90 (m, 2H),1.89-1.70 (m, 4H), 1.70-1.45 (m, 11H), 1.45-1.32 (m, 5H), 1.32-1.19 (m,9H), 1.19-1.08 (m, 3H), 1.08-0.98 (m, 5H), 0.98-0.89 (m, 4H), 0.84 (s,3H), 0.68 (s, 3H).

LCMS Rt=4.832 min in 7 min chromatography, 30-90AB_7MIN_E, purity 100%,MS ESI calcd. for C₂₉H₄₇ [M+H-2H₂O]⁺ 395, found 395.

HPLC Rt=6.338 min in 10 min chromatography, 50-100AB_10MIN.M, purity98%.

Example 59: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(59)

1. Pd(OH)₂ (200 mg, dry) was added to a solution of S-500-6-30 (140 mg,0.288 mmol) in MeOH (30 mL). The mixture was stirred at 50° C. under H₂(50 Psi) for 48 hrs. The mixture was filtered, concentrated and purifiedby combi-flash (0-15% of EtOAc in PE) to give S-500-6-25 (27 mg, 19%)and S-500-6-26 (42 mg, 30%) as a solid.

S-500-6-25:

¹H NMR (400 MHz, CDCl₃) δ 3.84-3.77 (m, 1H), 1.99-1.84 (m, 2H),1.81-1.72 (m, 1H), 1.68-1.56 (m, 4H), 1.53-1.43 (m, 5H), 1.42-1.32 (m,9H), 1.31-1.23 (m, 5H), 1.22-0.12 (m, 7H), 1.12-1.00 (m, 5H), 0.99-0.95(m, 4H), 0.94-0.85 (m, 12H), 0.66 (s, 3H).

LCMS Rt=1.797 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₅ [M+H-2H₂O]⁺ 451, found 451.

Example 60: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-((1R,2S)-2-methylcyclopropyl)butan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(60)

1. To a solution of N-8-7_2 (1 g, 2.41 mmol) in THF (50 mL) was addedLiAlH₄ (914 mg, 24.1 mmol) at 0° C. The grey suspension was heated at66° C. for 18 hrs. The reaction mixture was cooled to 0° C., quenched byice-water (914 mg), then 15% w/w aqueous NaOH (914 mg), and water (2.74g). The mixture was filtered and the filtrate cake was washed with DCM(3×50 mL). The filtrate was concentrated to give a residue, which waspurified by flash chromatography twice (ethyl acetate 10% in PE) to giveN-8-7_3 (192 mg, 19%) and N-8-7_3A (397 mg, 39%) as an oil.

N-8-7_3:

¹H NMR (400 MHz, CDCl3) δ 5.59-5.36 (m, 2H), 3.69-3.61 (m, 1H),2.25-2.12 (m, 1H), 2.08-1.81 (m, 3H), 1.68 (d, J=10.0 Hz, 3H), 1.64-1.54(m, 9H), 1.53-1.15 (m, 11H), 1.14-0.92 (m, 5H), 0.92-0.85 (m, 5H), 0.83(s, 4H), 0.69-0.60 (m, 4H).

N-8-7_3A:

¹H NMR (400 MHz, CDCl₃) δ 5.62-5.37 (m, 2H), 3.62 (br d, J=10.0 Hz, 1H),2.20-2.06 (m, 1H), 1.99-1.61 (m, 6H), 1.61-1.44 (m, 11H), 1.43-1.18 (m,5H), 1.16-0.94 (m, 6H), 0.94-0.85 (m, 5H), 0.82 (s, 5H), 0.70-0.58 (m,6H).

2. To a solution of diethylzinc (1 M in toluene, 4.31 mL, 4.31 mmol) inDCM (15 ml) at 0° C. was added CH₂I₂ (2.31 g, 8.63 mmol) over a periodof 15 min at 0° C. The milky suspension was stirred for 10 min at 0° C.and a preformed solution of Charette ligand((4R,5R)-2-(tert-butyl)-N4,N4,N5,N5-tetramethyl-1,3,2-dioxaborolane-4,5-dicarboxamide)(233 mg, 0.8638 mmol) and N-8-7_3A (300 mg, 0.7199 mmol) in DCM (20 ml)was rapidly added via syringe, whereupon the reaction mixture turnedclear. The solution was allowed to reach 25° C. and stirred for 16 h atthis temperature. The reaction was then quenched by addition ofsaturated aqueous NH₄Cl (150 ml), the phases were separated and theaqueous phase was extracted with DCM (3×100 ml). The combined organicphase was washed with saturated aqueous NaHCO₃ (150 mL), saturatedaqueous Na₂S₂O₃ (150 mL), brine (100 mL), dried over Na₂SO₄, filteredand concentrated under reduced pressure and the residue was purified byflash column chromatography (11% of ethyl acetate in PE) to affordN-8-7_4A (140 mg, 45%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.81-3.72 (m, 1H), 1.97-1.90 (m, 1H),1.76-1.58 (m, 4H), 1.53-1.44 (m, 4H), 1.43-1.15 (m, 10H), 1.13-0.91 (m,8H), 0.90-0.78 (m, 12H), 0.66 (s, 3H), 0.54-0.34 (m, 4H), 0.32-0.22 (m,2H), 0.19-0.12 (m, 1H).

3. To a solution of N-8-7_4A (140 mg, 0.325 mmol) in pyridine (5 mL) wasadded benzoyl chloride (91.3 mg, 0.65 mmol) followed by DMAP (15.8 mg,0.13 mmol) at 25° C. The reaction mixture was stirred at 60° C. for 16hours. The reaction mixture was diluted with DCM (80 mL). The DCM phasewas washed with water (100 mL), 1.0 M HCl aqueous (2×100 mL), 10% NaHCO₃aqueous (2×100 mL), brine (100 mL), dried over Na₂SO₄, filtered andconcentrated under vacuum to give an oil, which was purified by flashcolumn (1% of ethyl acetate in PE) to give N-8-7_5A (180 mg, impure) asan oil, which was further purified by flash column (PE) to give N-8-7_5A(110 mg, 61%) as a solid.

LCMS Rt=1.439 min in 2 min chromatography, 5-95AB_220&254, purity 93%,MS ESI calcd. for C₃₆H₅₃O₂ [M−H₂O+H]⁺517.8, found 517.8.

SFC Peak 1: Rt=4.079 min and Peak 2: Rt=4.345 min in 10 minchromatography, AD_3_EtOH_DEA_5_40_25ML (“Chiralpak AD-3 150×4.6 mmI.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5%to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 minFlow rate: 2.5 mL/min Column temp.: 35° C.″).

4. N-8-7A_5 (110 mg, 0.206 mmol) was purified by SFC (column: AD (250mm*30 mm, 5 um)), gradient: 30-30% B (A=0.1% NH₃/H₂O, B=EtOH), flowrate: 60 mL/min) to give impure N-8-7A_6 (Peak 1, 54 mg, 50%) as asolid, and impure N-8-8A_1 (Peak 2, 23 mg, impure) as a solid.

SFC Rt=4.088 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

5. To a solution of N-8-7A_6 (54 mg, 0.101 mmol) in THF/MeOH (1.5 mL/1.5mL), was added KOH (45.2 mg, 0.807 mmol) in water (0.5 mL). The reactionmixture was stirred at 50° C. for 16 hours. To the mixture was added HCl(0.2 M, 50 mL). The suspension was extracted with DCM (2×60 mL). Thecombined organic phase was washed with 3% aqueous NaHCO₃ (80 mL), brine(80 mL), dried over Na₂SO₄, filtered and concentrated under vacuum togive a solid, which was purified by flash-chromatography (15% of ethylacetate in PE) to give 60 (21 mg, 48%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.80-3.75 (m, 1H), 1.97-1.91 (m, 1H),1.76-1.58 (m, 7H), 1.54-1.27 (m, 7H), 1.26-1.06 (m, 7H), 1.04 (d, J=6.0Hz, 4H), 0.95 (s, 3H), 0.90-0.84 (m, 8H), 0.82 (s, 4H), 0.66 (s, 3H),0.64-0.59 (m, 1H), 0.52-0.37 (m, 2H), 0.32-0.22 (m, 2H).

LCMS Rt=1.327 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₉H₄₉O [M−H₂O+H]⁺413.4, found 413.4.

Example 61: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(61)

1. DMP (1.22 g, 2.88 mmol) was added to a solution of S-500-6-11_1 (580mg, 1.44 mmol) in DCM (30 mL). After that, the reaction mixture wasstirred at 15° C. for 10 min. The reaction mixture was quenched withSaturated NaHCO₃ aqueous (50 mL) until pH of the aqueous layer becameabout 9. The mixture was filtered. The DCM layer was separated and theaqueous phase was extracted with DCM (20 mL). The combined organic phasewas washed with saturated Na₂S₂O₃ aqueous (3×40 mL), sat.NaHCO₃ (40 mL),brine (40 mL), dried over Na₂SO₄, filtered and concentrated to givecrude S-500-6-11_1A (550 mg, Crude) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 5.34-5.25 (m, 1H), 2.58-2.29 (m, 4H),2.08-1.90 (m, 3H), 1.78-1.56 (m, 9H), 1.54-1.35 (m, 6H), 1.31-1.21 (m,2H), 1.19-1.08 (m, 5H), 1.06-0.99 (m, 5H), 0.93-0.82 (m, 6H), 0.69 (s,3H).

2. NaBH₄ (1.39 g, 41.1 mmol) was added in five two-minute intervals to asolution of S-500-6-11A (550 mg, 1.37 mmol) in THF (4 mL) and MeOH (2mL). The mixture was stirred at 15° C. for 30 minutes. The mixture wasquenched with sat.NH₄Cl (20 mL) and extracted with EtOAc (3×6 mL). Thecombined organic phase was dried over Na₂SO₄, filtered, concentrated andpurified by combi-flash (0-15% of EtOAc in PE) to give impure 61 (120mg) as a white solid, which was further purified by combi-flash (0-15%of EtOAc in PE) again to give pure 61 (150 mg, 75%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.25 (m, 1H), 3.73-3.59 (m, 1H),2.44-2.29 (m, 1H), 2.08-1.92 (m, 3H), 1.76-1.57 (m, 6H), 1.54-1.26 (m,10H), 1.25-1.18 (m, 3H), 1.17-1.06 (m, 4H), 1.03 (s, 3H), 1.00-0.88 (m,8H), 0.87-0.82 (m, 3H), 0.69 (s, 3H).

LCMS Rt=1.345 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₇H₄₅O [M+H-H₂O]⁺ 385, found 385.

Example 62: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(62)

1. To a solution of S-500-6-1_1 (500 mg, 1.39 mmol) and CsF (105 mg, 695umol) in THF (5 mL) was added TMSCF₃ (493 mg, 3.47 mmol) at 0° C. Themixture was stirred at 25° C. for 1 hr and treated with TBAF.3H₂O (1.09g, 3.47 mmol). The mixture was stirred at 25° C. for 2 hrs, quenchedwith water (100 mL) and extracted with EtOAc (2×50 mL). The combinedorganic phase was washed with brine (100 mL), dried over anhydrousNa₂SO₄, filtered and concentrated in vacuum. The residue was purified bysilica gel chromatography (100-200 mesh silica gel, PE/EA=10/1) toafford S-500-6-1_2 (400 mg, 67%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 5.33-5.24 (m, 1H), 4.06-4.00 (m, 1H),2.38-2.35 (m, 1H), 2.08-1.82 (m, 6H), 1.77-1.69 (m, 1H), 1.62-1.20 (m,13H), 1.16-1.00 (m, 8H), 0.99-0.92 (m, 1H), 0.87-0.83 (m, 4H), 0.74-0.64(m, 3H).

2. 3.5 g of S-500-6-1_2 was separated by SFC (column: AD (250 mm*30 mm,5 um), gradient: 40-40% B (A=0.05% NH₃/H₂O, B=MeOH), flow rate: 200mL/min) to give pure 81 (1 g, 28%, Peak 1) and 62 (1871 mg, 53%, Peak 2)as a white solid.

62:

¹H NMR (400 MHz, CDCl3) δ 5.30-5.28 (m, 1H), 4.03-3.99 (m, 1H),2.38-2.34 (m, 1H), 2.10-1.83 (m, 6H), 1.78-1.55 (m, 5H), 1.52-1.32 (m,6H), 1.31-1.01 (m, 12H), 0.98-0.92 (s, 1H), 0.85 (t, J=8 Hz, 3H), 0.73(s, 3H).

LCMS Rt=1.219 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₃₈F₃O [M+H-H₂O]⁺ 411, found 411.

SFC Peak 2: Rt=5.262 min in 10 min chromatography,AD_3_EtOH_DEA_5_40_25ML, 99% de.

Example 63: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-(tetrahydro-2H-pyran-4-yl)butan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(63)

1. Water (1 mL) and KOH (177 mg, 3.17 mmol) were added to a solution ofN-6-10_1 (180 mg, 0.318 mmol) in THF (2 mL) and methanol (1 mL). Themixture was stirred at 50° C. for 18 hrs. The reaction mixture wascooled, diluted with water (5 mL), acidified with 10% HCl (0.2 mL) andextracted with EtOAc (3×5 mL). The combined organic layer was dried overanhydrous sodium sulfate, filtered and concentrated. The residue waspurified by column chromatography on silica gel (10˜30% of EtOAc in PE)to give 63 (108 mg, 74%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.98-3.90 (m, 2H), 3.85-3.77 (m, 1H),3.44-3.33 (m, 2H), 1.99-1.82 (m, 2H), 1.69-1.58 (m, 6H), 1.57-1.45 (m,6H), 1.43-1.29 (m, 7H), 1.28-1.14 (m, 7H), 1.13-0.95 (m, 5H), 0.93-0.84(m, 7H), 0.83 (s, 3H), 0.69-0.61 (m, 4H).

LCMS Rt=1.167 min in 2.0 min chromatography, 30-90AB, purity 100%.

MS ESI calcd. for C₃₀H₄₉O [M-2H₂O+H]⁺ 425, found 425.

Example 64: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(64)

1. EtMgBr (42 mL, 126 mmol, 3 M in ether) was added slowly to a solutionof E-3_1 (20.0 g, 63.1 mmol) in THF (300 mL) under N₂ at −70° C. Afteraddition, the mixture was stirred at −70° C. for 2 hrs. The mixture wasquenched with sat. NH₄Cl (500 mL) and extracted with EtOAc (3×500 mL).The combined organic phase was washed with brine (500 mL), dried overNa₂SO₄, filtered, concentrated and purified by combi-flash (0-15% ofEtOAc in PE) to give E-3_2 (6.50 g, 30%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.57-2.49 (m, 1H), 2.23-1.80 (m, 6H),1.78-1.52 (m, 4H), 1.50-1.02 (m, 17H), 0.97 (s, 3H), 0.95-0.80 (m, 4H),0.60 (s, 3H).

2. t-BuOK (4.19 g, 37.4 mmol) was added to a suspension of MePPh₃Br(13.3 g, 37.4 mmol) in THF (200 mL) at 15° C. under N₂. The mixture wasstirred at 50° C. for 30 mins. To the mixture was added E-3_2 (6.50 g,18.7 mmol) in portions below 50° C. The mixture was stirred at 50° C.for 1 hr. To the mixture was added NH₄Cl (400 mL). The organic layer wasseparated and concentrated under vacuum to give a crude product, whichwas triturated from MeOH/water (200 mL, 1:1) at 50° C. The mixture wasfiltered after cooled and the solid was washed with MeOH/water (2×30 mL,1:1) and concentrated in vacuum to give E-3_3 (5.8 g, impure) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.06-1.97 (m, 1H),1.94-1.55 (m, 12H), 1.53-1.05 (m, 16H), 0.97 (s, 3H), 0.95-0.85 (m, 3H),0.55 (s, 3H). 3. 9-BBN dimer (8.19 g, 33.6 mmol) was added to a mixtureof E-3_3 (5.80 g, 16.8 mmol) in THF (100 mL) at 15° C. under N₂. Thereaction mixture was stirred at 60° C. for 1 hour. The mixture wascooled to 15° C. Ethanol (7.72 g, 168 mmol) was added at 15° C. NaOHaqueous (33.6 mL, 5 M, 168 mmol) was added dropwise at 15° C. H₂O₂ (16.8mL, 10.0 M, 168 mmol) was added dropwise at 15° C. The obtained mixturewas stirred at 60° C. for 1 hour. The aqueous phase was extracted withEtOAc (3×100 mL). The combined organic phase was washed with brine(2×100 mL), dried over Na₂SO₄, filtered and concentrated. The residuewas triturated from CH₃OH/H₂O=1/1 (150 mL) at 65° C. to give E-3_4 (2.80g, 46%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.61 (m, 1H), 3.38-3.32 (m, 1H),2.03-1.56 (m, 4H), 1.56-1.51 (m, 5H), 1.51-1.10 (m, 16H), 1.10-1.02 (m,6H), 0.97 (s, 3H), 0.96-0.88 (m, 3H), 0.67 (s, 3H).

4. DMP (5.80 g, 13.7 mmol) was added to a solution of E-3_4 (2.50 g,6.89 mmol) in DCM (50 mL). After that, the reaction was stirred at 20°C. for 30 min. The reaction mixture was added saturated aqueous NaHCO₃(50 mL) solution, aqueous saturated Na₂S₂O₃ (30 mL) solution, extractedwith DCM (2×20 mL). The combined organic layer was washed with aqueoussaturated NaHCO₃ (3×10 mL) solution and brine (20 mL), dried overNa₂SO₄, filtered and concentrated under vacuum to give E-3_5 (2.45 g,crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.60-9.55 (m, 1H), 2.26 (s, 1H), 1.98-1.70 (m,6H), 1.70-1.51 (m, 6H), 1.51-1.00 (m, 12H), 1.00-0.89 (m, 10H),1.75-0.65 (m, 4H).

5. Isobutylmagnesium bromide (33.9 mL, 2 M in THF, 67.9 mmol) was addedto a solution of E-3_5 (2.45 g, 6.79 mmol) in THF (10 mL) under N₂ at 0°C. The mixture was stirred at 20° C. for 16 hrs. To the mixture wasadded NH₄Cl (20 mL, sat. aq.), the mixture was extracted with EtOAc(2×30 mL). The combined organic phase was washed with brine (20 mL),dried over Na₂SO₄, concentrated under vacuum, and purified by flashcolumn (0˜20% of EtOAc in PE) to give E-3_6 (1.6 g, 55%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.65-3.55 (m, 1H), 2.01-1.85 (m, 3H),1.85-1.49 (m, 3H), 1.49-1.36 (m, 12H), 1.36-1.22 (m, 10H), 1.22-1.02 (m,9H), 1.02-0.98 (m, 4H), 0.98-0.80 (m, 7H), 0.66 (s, 3H).

6. DMP (3.12 g, 7.38 mmol) was added to a solution of E-3_6 (1.6 g, 3.69mmol) in DCM (30 mL). After that, the reaction was stirred at 20° C. for30 min. To the reaction mixture was added saturated aqueous NaHCO₃ (20mL) solution, aqueous saturated Na₂S₂O₃ (20 mL) solution, followed byextraction with DCM (2×20 mL). The combined organic layer was washedwith aqueous saturated NaHCO₃ (3×10 mL) solution and brine (20 mL),dried over Na₂SO₄, filtered and concentrated under vacuum to give E-3_7(1.5 g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 2.55-2.80 (m, 3H), 2.80-2.23 (m, 1H),1.98-1.81 (m, 2H), 1.81-1.1.69 (m, 1H), 1.69-1.25 (m, 16H), 1.25-1.01(m, 10H), 1.01-0.81 (m, 14H), 0.67 (s, 3H).

7. NaBH₄ (255 mg, 6.72 mmol) was added in one portion to a solution ofE-3_7 (1.45 g, 3.36 mmol) in MeOH (20 mL) at 0° C. After addition, themixture was stirred at 20° C. for 1 hr and quenched with NH₄Cl (20 mL,sat. aq.). The mixture was extracted with DCM (2×20 mL). The combinedorganic phase was washed with brine (2×10 mL), dried over Na₂SO₄,filtered, concentrated and purified by flash column (0˜20% of EtOAc inPE) to give E-3_8 (1.2 g, 83%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.80-3.70 (m, 1H), 3.70-3.55 (m, 1H),2.05-1.82 (m, 3H), 1.82-1.55 (m, 4H), 1.55-1.35 (m, 5H), 1.35-1.00 (m,18H), 1.00-0.79 (m, 17H), 0.66 (s, 3H).

8. Benzoyl chloride (1.85 g, 13.2 mmol) was added to a solution of E-3_8(1.15 g, 2.65 mmol) in pyridine (20 mL). The reaction mixture wasstirred at 20° C. for 4 hours. The reaction mixture was poured intowater (20 mL). The mixture was extracted with EtOAc (2×20 mL). Thecombined organic phase was washed with saturated brine (2×10 mL), driedover anhydrous Na₂SO₄, filtered concentrated and purified by flashcolumn (0˜10% of EtOAc in PE) to give the mixture product (1.45 g,impure) as a solid. The mixture product (1.45 g, impure) was purified bySFC (column: AD (250 mm*50 mm, 10 um), gradient: 30-30% B (A=0.1%NH3/H2O, B=EtOH), flow rate: 200 mL/min) to give E-3_9 (peak 2, 470 mg,33%, DE %=100%) as a solid and E-3_9A (peak 1, 600 mg, 42%, DE %=99.1%)as a solid.

E-3_9A:

¹H NMR (400 MHz, CDCl₃) δ 8.10-7.99 (m, 2H), 7.60-7.50 (m, 1H),7.50-7.38 (m, 2H), 5.15-5.05 (m, 1H), 2.05-1.70 (m, 6H), 1.70-1.35 (m,5H), 1.35-1.05 (m, 19H), 1.05-0.82 (m, 17H), 0.65 (s, 3H).

SFC Rt=3.344 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de. E-3_9:

¹H NMR (400 MHz, CDCl₃) δ 8.10-7.99 (m, 2H), 7.60-7.50 (m, 1H),7.50-7.38 (m, 2H), 5.25-5.15 (m, 1H), 2.05-1.80 (m, 3H), 1.80-1.45 (m,15H), 1.45-1.09 (m, 13H), 1.09-0.85 (m, 16H), 0.68 (s, 3H).

SFC Rt=3.851 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML,99.1% de.

9. NaOH (531 mg, 13.3 mmol) and H₂O (0.5 mL) were added to a solution ofE-3_9A (600 mg, 1.11 mmol) in THF (2 mL) and MeOH (2 mL) at 25° C. Thesolution was stirred at 50° C. for 48 hrs. Water (10 mL) was added. Themixture was extracted with EtOAc (2×10 mL). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated in vacuum to givecrude product which was triturated with MeCN (10 mL) to give desiredproduct 69 (473 mg, 99%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.68-3.55 (m, 1H), 2.01-1.85 (m, 3H),1.85-1.70 (m, 1H), 1.70-1.45 (m, 8H), 1.45-1.22 (m, 13H), 1.22-1.05 (m,8H), 1.05-1.86 (m, 15H), 0.66 (s, 3H).

LCMS t_(R)=1.403 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₉H₄₉ [M+H-2H₂O]⁺ 397, found 397.

Example 65: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-methylpentan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(65)

N-8-11_1 (30 mg, 0.0741 mmol, impure) was purified by combi-flash (25%of EtOAc in PE) to give 65 (9 mg, 30%) as a solid.

65:

¹H NMR (400 MHz, CDCl₃) δ 3.18-3.07 (m, 1H), 1.98-1.81 (m, 2H),1.71-1.58 (m, 6H), 1.53-1.31 (m, 7H), 1.30-0.98 (m, 14H), 0.97-0.78 (m,14H), 0.70-0.60 (m, 4H).

LCMS Rt=4.387 min in 7.0 min chromatography, 30-90AB_7MIN_E, purity97.6%, MS ESI calcd for C₂₇H₄₅ [M+H-2H₂O]⁺ 369, found 369.

Example 66: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-(2-methylcyclopropyl)butan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(66)

1. To a solution of N-8-7_2 (1 g, 2.41 mmol) in THF (50 mL) was addedLiAlH₄ (914 mg, 24.1 mmol) at 0° C. The grey suspension was heated at66° C. for 18 hrs. The reaction mixture was cooled to 0° C., quenched byice-water (914 mg), then 15% w/w NaOH aqueous (914 mg), water (2.74 g).The mixture was filtered and the filtrate cake was washed with DCM (3×50mL). The filtrate was concentrated to give a residue, which was purifiedby flash chromatography twice (ethyl acetate 10% in PE) to give N-8-7_3(192 mg, 19%) and N-8-7_3A (397 mg, 39%) as an oil.

N-8-7_3:

1H NMR (400 MHz, CDCl3) δ 5.59-5.36 (m, 2H), 3.69-3.61 (m, 1H),2.25-2.12 (m, 1H), 2.08-1.81 (m, 3H), 1.68 (d, J=10.0 Hz, 3H), 1.64-1.54(m, 9H), 1.53-1.15 (m, 11H), 1.14-0.92 (m, 5H), 0.92-0.85 (m, 5H), 0.83(s, 4H), 0.69-0.60 (m, 4H).

N-8-7_3A:

¹H NMR (400 MHz, CDCl₃) δ 5.62-5.37 (m, 2H), 3.62 (br d, J=10.0 Hz, 1H),2.20-2.06 (m, 1H), 1.99-1.61 (m, 6H), 1.61-1.44 (m, 11H), 1.43-1.18 (m,5H), 1.16-0.94 (m, 6H), 0.94-0.85 (m, 5H), 0.82 (s, 5H), 0.70-0.58 (m,6H).

2. To a solution of diethylzinc (1 M in toluene, 2.59 mL, 2.59 mmol) inDCM (10 ml) was added CH₂I₂ (1.38 g, 5.18 mmol) over a period of 15 minat 0° C. The milky suspension was stirred for 10 min at 0 C and apreformed solution of Charette ligand((4R,5R)-2-(tert-butyl)-N4,N4,N5,N5-tetramethyl-1,3,2-dioxaborolane-4,5-dicarboxamide)(139 mg, 0.5182 mmol) and N-8-7_3 (180 mg, 0.4319 mmol) in DCM (15 ml)was rapidly added via syringe, whereupon the reaction mixture turnedclear. The solution was allowed to be warmed up to 25° C. and stirredfor 16 h at 25° C. The reaction was then quenched by addition ofsaturated aqueous NH₄Cl (150 ml). The phases were separated and theaqueous phase was extracted with DCM (3×60 ml). The combined organicphase was washed with saturated NaHCO₃ aqueous (150 mL), saturatedaqueous Na₂S₂O₃ (150 mL), and brine (100 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure and the residue waspurified by flash column chromatography (11% of ethyl acetate in PE) toafford N-8-7_4 (60 mg, 32%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.77 (br s, 1H), 1.99-1.87 (m, 2H), 1.70-1.56(m, 6H), 1.54-1.34 (m, 6H), 1.31-1.16 (m, 5H), 1.15-1.05 (m, 1H),1.15-0.97 (m, 7H), 0.99-0.94 (m, 1H), 0.92-0.77 (m, 10H), 0.69-0.58 (m,6H), 0.52-0.13 (m, 5H).

3. To a solution of N-8-7_4A (60 mg, 0.1393 mmol) in pyridine (3 mL) wasadded benzoyl chloride (39.1 mg, 0.2786 mmol) followed by DMAP (6.79 mg,0.05572 mmol) at 25° C. The reaction mixture was stirred at 60° C. for16 hours. The reaction mixture was diluted with DCM (80 mL). The DCMphase was washed with water (100 mL), 1.0 M HCl aqueous (2×100 mL), 10%NaHCO₃ aqueous (2×100 mL), brine (100 mL), dried over Na₂SO₄, filteredand concentrated under vacuum to give an oil, which was purified byflash column (1% of ethyl acetate in PE) to give N-8-7_5 (24 mg, 32%) asan oil.

LCMS Rt=1.431 min in 2 min chromatography, 5-95AB_220&254, purity 90%,MS ESI calcd. for C₃₆H₅₃O₂ [M−H₂O+H]⁺517.3, found 517.3.

SFC Peak 1: Rt=5.703 min in 10 min chromatography,AD_3_EtOH_DEA_5_40_25ML (“Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobilephase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5mL/min Column temp.: 35° C.″).

4. N-8-7_5 (24 mg, 0.04487 mmol) was purified by SFC (column: AD (250mm*30 mm, 5 um)), gradient: 40-40% B (A=0.1% NH₃/H₂O, B=EtOH), flowrate: 50 mL/min) to give impure N-8-7_6 (RT: 5.732, 19 mg, impure) as asolid. No isomer was obtained.

LCMS Rt=1.435 min in 2 min chromatography, 5-95AB_220&254, purity 98%,MS ESI calcd. for C₃₆H₅₃O₂ [M−H₂O+H]⁺517.3, found 517.3.

SFC Rt=5.732 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML,97.76% de.

5. To a solution of N-8-7_6 (19 mg, 0.0355 mmol) in THF/MeOH (0.5 mL/0.5mL), was added KOH (19.8 mg, 0.0.3552 mmol) in water (0.2 mL). Thereaction mixture was stirred at 55° C. for 16 hours. To the mixture wasadded HCl (0.2 M, 50 mL). The suspension was extracted with DCM (2×60mL). The combined organic phase was washed with 3% NaHCO₃ aqueous (80mL) and brine (80 mL), dried over Na₂SO₄, filtered and concentratedunder vacuum to give a solid, which was purified by flash-chromatography(ethyl acetate in PE, 15%) to give 66 (2 mg, 13%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.77 (br s, 1H), 1.98-1.87 (m, 2H), 1.70-1.57(m, 7H), 1.54-1.28 (m, 11H), 1.26-0.94 (m, 12H), 0.91-0.84 (m, 7H), 0.83(s, 3H), 0.72-0.60 (m, 4H), 0.51-0.15 (m, 3H).

LCMS Rt=1.315 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₉H₄₉O [M−H₂O+H]⁺395.3, found 395.3.

Example 67: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(67)

1. Pd(OH)₂ (160 mg, dry) was added to a solution of 32 (80 mg, 0.18mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 67 (10 mg, 12%) and 84 (30mg, 37%) as a solid.

67:

¹H NMR (400 MHz, CDCl₃) δ 3.76-3.66 (m, 1H), 2.01-1.78 (m, 5H),1.76-1.58 (m, 7H), 1.52-1.31 (m, 13H), 1.28-1.10 (m, 10H), 1.09-0.99 (m,4H), 0.96 (s, 3H), 0.93-0.86 (m, 6H), 0.67 (s, 3H).

LCMS Rt=1.508 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₀H₄₉ [M+H-2H₂O]⁺ 409, found 409.

Example 68: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxyhept-5-yn-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(68)

1. The crude N-8-7_2B (250 mg, 0.868 mmol) was further purified by SFC(column: AD (250 mm*30 mm, 10 um)), gradient: 35-35% B (A=0.1% NH₃/H₂O,B=EtOH), flow rate: 60 mL/min) to give 41 (peak 2, 81 mg, 33%) as asolid and 68 (peak 1, 78 mg, 31%) as a solid.

68:

¹H NMR (400 MHz, CDCl₃) δ 3.87-3.78 (m, 1H), 2.21-2.12 (m, 1H),1.99-1.86 (m, 2H), 1.80 (s, 3H), 1.73-1.51 (m, 8H), 1.51-1.42 (m, 4H),1.42-1.20 (m, 8H), 1.20-0.95 (m, 7H), 0.95-0.79 (m, 8H), 0.95 (s, 4H).

LCMS Rt=1.188 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₈H₄₅O [M+H-H₂O]⁺ 397 found 397.

SFC Rt=6.465 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

Example 69: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3S)-5-cyclopropyl-3-hydroxypentan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(69)

1. NaBH₄ (2.46 g, 65.1 mmol) was added in portions to a solution ofS-500-6-19_3 (700 mg, 1.63 mmol) in THF (5 mL) and MeOH (5 mL) at 15° C.After stirring at 15° C. for 20 mins, the mixture was quenched withNH₄Cl (20 mL, sat. aq.) and extracted with EtOAc (50 mL). The organiclayer was separated and concentrated in vacuum to give 760 mg mixture asa solid, which was separated by flash column (0˜35% of DCM/EtOAc (1/1)in PE) to give 69 (330 mg, 47%) and 6 (250 mg, 35%, impure) as a solid.The impure 6 (250 mg) was further separated by flash column (0˜35% ofDCM/EtOAc (1/1) in PE) to give 6 (170 mg, 23%) as a solid.

69:

¹H NMR (400 MHz, CDCl₃) δ 5.33-5.23 (m, 1H), 3.75-3.63 (m, 1H),2.41-2.31 (m, 1H), 2.09-1.85 (m, 4H), 1.78-1.59 (m, 5H), 1.53-1.38 (m,9H), 1.38-1.05 (m, 9H), 1.03 (s, 3H), 1.00-0.91 (m, 1H), 0.91 (d, J=6.4Hz, 3H) 0.85 (t, J=7.6 Hz, 3H), 0.69 (s, 3H), 0.68-0.60 (m, 1H),0.45-0.36 (m, 2H), 0.09-0.08 (m, 2H).

LCMS Rt=1.387 min in 2.0 min chromatography, 30-90_AB_E, purity 98.1%,MS ESI calcd. for C₂₉H₄₇O [M+H-H₂O]⁺ 411, found 411.

S-500-6-19:

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.24 (m, 1H), 3.77-3.66 (m, 1H),2.41-2.31 (m, 1H), 2.09-1.91 (m, 3H), 1.79-1.59 (m, 6H), 1.55-1.21 (m,14H), 1.21-1.06 (m, 4H), 1.03 (s, 3H), 1.00-0.95 (m, 1H), 0.93 (d, J=6.8Hz, 3H) 0.85 (t, J=7.6 Hz, 3H), 0.70 (s, 3H), 0.68-0.62 (m, 1H),0.49-0.38 (m, 2H), 0.11-0.02 (m, 2H).

LCMS Rt=1.380 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₉H₄₇O [M+H-H₂O]⁺ 411, found 411.

Example 70: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-4-(3-methyloxetan-3-yl)butan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(70)

1. To a suspension of Mg (807 mg, 33.2 mmol) and I₂ (1 mg) in THF (2 mL)was added solution of N-014-005_3 (2.5 g, 15.1 mmol) in THF (8 mL) dropwise under N₂ at 50˜55° C. The mixture was stirred at 55° C. for 1 h.The mixture was diluted with THF (10 mL) and used in the next stepdirectly without monitored. To a solution of N-14-12_4 (1.01 g, 2.83mmol) in THF (10 mL) was added fresh prepared 3-[(bromomagnesio)methyl]-3-methyloxetane (15 mmol in 20 mL of THF) at 0° C. The mixturewas stirred at 15° C. for 4 h. To the mixture was added NH₄Cl (20 mL,10% aq.). The mixture was extracted with EtOAc (30 mL). The organiclayer was separated and concentrated in vacuum. The residue was purifiedby flash column (0˜30% of EtOAc in PE) to give a mixture (190 mg, 15%)as a white solid, which was purified by SFC (Column: AD (250 mm*30 mm, 5um), Condition: 0.1% NH₃H₂O ETOH, Gradient: from 50% to 50%, FlowRate(ml/min): 60 mL/min, 25° C.) to afford 33 (Peak 1, 110 mg, 9%) and 70(Peak 2, 30 mg, impure) as a white solid. The impure 70 (30 mg, impure)was purified by column chromatography on silica gel (15% of EtOAc in PE)to give 70 (10 mg, 5%) as a white solid.

33:

¹H NMR (400 MHz, CDCl₃) δ 5.30-5.26 (m, 1H), 4.59-4.70 (m, 1H),4.50-4.48 (m, 1H), 4.36-4.33 (m, 1H), 3.83 (s, 1H), 2.40-2.33 (m, 1H),2.10-1.50 (m, 17H), 1.49-1.35 (m, 9H), 1.30-0.80 (m, 13H), 0.68 (s, 3H).

LCMS Rt=1.069 min in 3 min chromatography, 30-90AB_2MIN_E.M, purity100%, MS ESI calcd. for C₂₉H₄₉O₃ [M+H]⁺ 445, found 445.

70:

¹H NMR (400 MHz, CDCl₃) δ 5.30-5.26 (m, 1H), 4.10-4.02 (m, 1H),3.81-3.70 (m, 1H), 3.50-3.41 (m, 2H), 3.34-3.30 (m, 1H), 2.35-2.31 (m,1H), 2.10-1.50 (m, 18H), 1.49-1.05 (m, 13H), 1.05-0.90 (m, 4H),0.90-0.80 (m, 3H), 0.67 (s, 3H).

LCMS Rt=1.115 min in 3 min chromatography, 30-90AB_2MIN_E.M, purity100%, MS ESI calcd. for C₂₉H₄₉O₃ [M+H]⁺ 445, found 445.

Example 71: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-(methoxymethyl)-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(71)

1. To a solution of N-004-022_5 (200 mg, 0.531 mmol), CsF (40.2 mg,0.265 mmol) in THF (5 mL) was added TMSCF₃ (187 mg, 1.32 mmol) under N₂at 0° C. The mixture was stirred at 25° C. for 1 hrs. To the mixture wasadded TBAF.3H₂O (836 mg, 2.65 mmol). After stirring at 25° C. for 2 hrs,the mixture was quenched 50% NH₄Cl (20 mL) and extracted with EtOAc(2×10 mL). The combined organic phase was washed with brine (20 mL),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. Theresidue was purified by silica gel chromatography (100-200 mesh silicagel, PE/EA=10/1) to afford 9 (56 mg, 24%) and 71 (30 mg, impure) as awhite solid.

71 (30 mg, 0.067 mmol) was re-crystallized from n-hexane (2 mL) at 25°C. to give 71 (24 mg, 10%) as a white solid.

71:

¹H NMR (400 MHz, CDCl₃) δ 4.08-4.00 (m, 1H), 3.39 (s, 3H), 3.24-3.18 (m,2H), 2.22-2.15 (m, 1H), 2.02-1.77 (m, 5H), 1.75-1.68 (m, 2H), 1.64-1.52(m, 5H), 1.47-1.31 (m, 6H), 1.28-1.01 (m, 10H), 0.97 (s, 3H), 0.67 (s,3H).

LCMS Rt=1.105 min in 2 min chromatography, 30-90AB_POS_E.M, purity 100%,MS ESI calcd. for C₂₅H₄₁F₃O₃ [M+Na]⁺ 469, found 469.

Example 72: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxyhexan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(72)

1. Propylmagnesium bromide (3.34 mL, 6.69 mmol, 2M in THF) was slowlyadded to a solution of S-500-6-1_1 (800 mg, 2.23 mmol) in THF (30 mL) at0° C. After addition, the mixture was stirred at 15° C. for 1 hr. Themixture was quenched with sat.NH₄Cl (40 mL) and extracted with EtOAc(3×20 mL). The combined organic phase was washed with brine (2×30 mL),dried over Na₂SO₄, filtered and concentrated and purified by combi-flash(0-15% of EtOAc in PE) to give 72 (500 mg, 56%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.31-5.26 (m, 1H), 3.72-3.64 (m, 1H),2.41-2.31 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.69 (m, 1H), 1.62-1.54 (m,3H), 1.52-1.38 (m, 9H), 1.37-1.16 (m, 6H), 1.15-1.01 (m, 7H), 0.99-0.88(m, 7H), 0.87-0.82 (m, 3H), 0.68 (s, 3H).

LCMS Rt=4.979 min in 7.0 min chromatography, 30-90AB_E, purity 98.8%, MSESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

Example 73: Synthesis of(3S,5S,8R,9R,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(73)

1. To solution of G-21-1 (40 g, 110 mmol) in dry dioxane (1 L) under N2was added sodium methoxide (29.7 g, 550 mmol). The mixture was stirredat 110° C. for 16 hours. TLC showed the starting material was consumedalmostly. Solvent was removed to ⅓ volume and mixture was acidified with2 M HCl to pH=5˜6, extracted with DCM (500 mL*3), washed with aqueoussodium bicarbonate (500 mL) and brine (500 mL), dried over sodiumsulfate and concentrated. The residue was purified by columnchromatography on silica gel (PE:EA:MeOH=3:1:0.1) to give G-21-4A (11 g,33.1%) as white solid.

2. To a solution of Li (2.54 g, 363 mmol) was added to liquid ammonium(1000 mL, prepared in 13-601 over 1.5 hours) at −70° C. in portions. Themixture was stirred at −70° C. for 30 mins until all of Li wasdissolved. A solution of G-21-4A (11 g, 36.3 mmol) and tert-BuOH (5.38g, 72.6 mmol) in 400 ml of anhydrous tetrahydrofuran was added dropwiseand stirred for 90 mins until the reaction mixture turned light yellow.TLC (PE:EA=1:1, PMA) showed most of STM was consumed. Ammonium chloride(15 g) was added and excess ammonia was left to evaporate. The residuewas extracted with 0.5N HCl (500 mL) and dichloromethane (500 mL×2). Thecombined organic layers were washed with saturated NaHCO₃ solution,dried over Na₂SO₄, filtered and concentrated to give a mixture ofG-21-5A and G-21-5B (10 g, impure) which was directly used in the nextstep without further purification.

3. A solution of G-21-5A and G-21-5B (10 g, 27.9 mmol) in 100 mL ofanhydrous dichloromethane was added PCC (16.6 g, 65.6 mmol) and silicongel (16.6 g). After stirring at 25° C. for 2 h, TLC (PE:EA=1:1, PMA)showed the STM was consumed. The resulting solution was concentrated andpurified by chromatography on silicon gel (petroleum ether/ethylacetate=5:1 to 2:1) to afford G-21-6A (4.6 g, 46.4%) as a white solid.

4. To a solution of BHT (34.8 g, 158 mmol) in toluene (120 mL) undernitrogen at 0° C. was added trimethylaluminum (2 M in toluene, 39.5 mL,79.1 mmol) dropwise. After stirring at 20° C. for 30 minutes, a solutionof G-21-6A (8 g, 26.4 mmol) in toluene (80 mL) was added dropwise undernitrogen at −70° C. After stirring at −70° C. for 30 min, MeMgBr (3 M indiethyl ether, 26.3 mL, 79.1 mmol, 3M in ether) was added dropwise. Theresulting mixture was stirred at −70° C. for 1 hr, poured to ice-cooledaqueous citric acid (300 mL) slowly and extracted with EtOAc (3×100 mL).The combined organic layer was washed with brine (300 mL), dried overanhydrous sodium sulfate, filtered, concentrated and purified bycombi-flash (0-40% of EtOAc in PE) to give N-4-10_1 (6.5 g, 77%) as awhite solid.

¹H NMR (400 MHz, CDCl₃) δ 2.60-2.49 (m, 1H), 2.47-2.37 (m, 2H),2.34-2.19 (m, 3H), 2.14-2.03 (m, 1H), 2.00-1.82 (m, 3H), 1.73-1.58 (m,3H), 1.56-1.46 (m, 2H), 1.36-1.26 (m, 3H), 1.24 (s, 3H), 1.21-1.07 (m,2H), 1.04 (s, 3H), 1.00-0.84 (m, 1H), 0.82 (s, 3H).

5. To a suspension of bromo(ethyl)triphenylphosphorane (22.6 g, 61.1mmol) in anhydrous THF (200 mL) under N₂ at 20° C. was added potassium2-methylpropan-2-olate (6.84 g, 61.1 mmol). After stirring at 40° C. for30 minutes, a solution of N-4-10_1 (6.5 g, 20.4 mmol) in anhydrous THF(50 mL) was added slowly. The resultant mixture was stirred at 40° C.for 10 minutes, then quenched with aqueous NH₄Cl (400 mL) and extractedwith EtOAc (2×150 mL). The combined organic phase was dried over sodiumsulfate, filtered, concentrated and purified by column chromatography onsilica gel (0-25% of EtOAc in PE) to give N-4-10_2 (5.5 g, 82%) as awhite solid.

¹H NMR (400 MHz, CDCl₃) δ 5.22-5.13 (m, 1H), 2.91-2.81 (m, 1H),2.62-2.51 (m, 1H), 2.50-2.39 (m, 2H), 2.38-2.24 (m, 1H), 1.91-1.81 (m,1H), 1.80-1.70 (m, 4H), 1.55-1.41 (m, 4H), 1.36-1.25 (m, 5H), 1.23 (s,3H), 1.21-1.04 (m, 3H), 1.01 (s, 3H), 0.98-0.84 (m, 2H), 0.81 (s, 3H).

6. To a mixture of N-4-10_2 (5.5 g, 16.6 mmol) in THF (100 mL) was added9-BBN dimer (8.10 g, 33.2 mmol) at 15° C. under N₂. After stirring at50° C. for 1 hour, the mixture was cooled to 15° C. NaOH aqueous (33.2mL, 5 M, 166 mmol) was added dropwise below 15° C., followed by adropwise addition of H₂O₂ (18.8 g, 30%, 166 mmol) below 15° C. Themixture was extracted with EtOAc (3×100 mL). The combined organic phasewas washed with sat.Na₂S₂O₃ (5×100 mL), dried over Na₂SO₄, filtered andconcentrated to give 7 g crude, which was used in next step directly.

7. To a solution of N-4-10_3 (7 g, 19.9 mmol) in DCM (300 mL) was addedDMP (25.2 g, 59.6 mmol). After stirring at 20° C. for 10 min, thereaction mixture was quenched with saturated NaHCO₃ solution (500 mL)until pH of the aqueous layer became about 9. The mixture was filtered.The DCM layer was separated and the aqueous phase was extracted with DCM(200 mL). The combined organic phase was washed with saturated Na₂S₂O₃aqueous (3×400 mL), sat.NaHCO₃ (400 mL), brine (400 mL), dried overNa₂SO₄, filtered, concentrated and purified by combi-flash (0-20% ofEtOAc in DCM) to give N-4-10_4 (4 g, 58%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 2.77-2.67 (m, 1H), 2.65-2.38 (m, 3H),2.32-2.17 (m, 1H), 2.09 (s, 3H), 1.88-1.63 (m, 7H), 1.59-1.49 (m, 3H),1.35-1.21 (m, 7H), 1.19-1.09 (m, 2H), 1.01 (s, 3H), 0.96-0.84 (m, 1H),0.57 (s, 3H).

8. To a suspension of MePh₃PBr (8.18 g, 23.0 mmol) in THF (100 mL) wasadded t-BuOK (2.57 g, 23.0 mmol). After stirring at 40° C. for 10minutes, the mixture was slowly added dropwise to a solution of N-4-10_4(4 g, 11.5 mmol) in THF (50 mL) at 20° C. After addition, the mixturewas quenched with NH₄Cl (200 mL) and extracted with EtOAc (3×80 mL). Thecombined organic phase was dried over Na₂SO₄, filtered, concentrated andpurified by combi-flash (0-25% of EtOAc in PE) to give N-4-10_5 (3.2 g,80%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 4.88 (s, 1H), 4.70 (s, 1H), 2.48-2.37 (m, 2H),2.31-2.22 (m, 2H), 1.89-1.77 (m, 4H), 1.75-1.61 (m, 7H), 1.54-1.45 (m,2H), 1.34-1.29 (m, 2H), 1.28-1.24 (m, 3H), 1.23 (s, 3H), 1.17-1.05 (m,2H), 1.01 (s, 3H), 0.93-0.83 (m, 1H), 0.51 (s, 3H).

9. To a mixture of N-4-10_5 (3.2 g, 9.28 mmol) in THF (100 mL) was added9-BBN dimer (4.51 g, 18.5 mmol) at 15° C. under N₂. After stirring at50° C. for 1 hour, the mixture was cooled to 15° C. NaOH aqueous (18.5mL, 5 M, 92.8 mmol) was added dropwise below 15° C., followed by adropwise addition of H₂O₂ (10.5 g, 30%, 92.8 mmol) below 15° C. Themixture was extracted with EtOAc (3×100 mL). The combined organic phasewas washed with sat.Na₂S₂O₃ (5×100 mL), dried over Na₂SO₄, filtered andconcentrated to give 5 g crude, which was used in next step directly.

10. To a solution of N-4-10_5A (5 g, 13.7 mmol) in DCM (300 mL) wasadded DMP (11.6 g, 27.4 mmol). After stirring at 20° C. for 10 min, thereaction mixture was quenched with saturated NaHCO₃ solution (300 mL)until pH of the aqueous layer became about 9. The mixture was filtered.The DCM layer was separated and the aqueous phase was extracted with DCM(100 mL). The combined organic phase was washed with saturated Na₂S₂O₃aqueous (3×300 mL), sat.NaHCO₃ (300 mL), brine (300 mL), dried overNa₂SO₄, filtered, concentrated and purified by combi-flash (0-10% ofAcetone in DCM) to give N-4-10_7 (1 g, 20%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 9.58-9.55 (m, 1H), 2.52-2.27 (m, 4H),2.08-1.96 (m, 1H), 1.84-1.62 (m, 8H), 1.51-1.39 (m, 3H), 1.32-1.21 (m,7H), 1.17-1.06 (m, 5H), 1.01 (s, 3H), 0.94-0.83 (m, 1H), 0.66 (s, 3H).

11. To a solution of N-4-10_6 (400 mg, 0.832 mmol) in THF (20 mL) wasadded dropwise isopentylmagnesium bromide (1.65 mL, 3.30 mmol, 2M inether) under N₂ at 0° C. After stirring at 0° C. for 10 minutes, themixture was quenched with sat.NH₄Cl (60 mL) and extracted with EtOAc(2×30 mL). The combined organic phase was washed with brine (60 mL),dried over Na₂SO₄, filtered, concentrated and purified by combi-flash(0-10% of Acetone in DCM) to give 73 (200 mg, 42%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 3.65-3.56 (m, 1H), 2.55-2.49 (m, 1H),2.46-2.38 (m, 1H), 2.32-2.25 (m, 1H), 2.10-1.98 (m, 1H), 1.83-1.62 (m,7H), 1.57-1.44 (m, 4H), 1.42-1.25 (m, 7H), 1.24-1.20 (m, 4H), 1.19-1.04(m, 5H), 1.01 (s, 3H), 0.94-0.82 (m, 10H), 0.0.63 (s, 3H).

LCMS Rt=3.381 min in 7.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₈H₄₇O₂ [M+H-H₂O]⁺ 415, found 415.

Example 74: Synthesis of(3S,5S,8R,9R,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(74)

1. t-BuOH (1.7 L) was charged into a three-neck round bottom flask underN₂ at 35° C. and stirred for 10 mins. t-BuOK (292 g, 2.61 mol) was addedto the mixture and stirred until the reaction became clear. After that,S-310-B9_1 (65 g, 238 mmol) was added to the above mixture and stirredfor 1.5 h at 35° C. under N₂. The reaction mixture was poured into 10%aqueous acetic acid (2 L) and stirred for 30 mins, during which thetemperature was maintained below 10° C. Then the mixture was treatedwith water (1.5 L) and the pH was adjusted to 7-8 with NaHCO₃ and themixture was stirred for 30 mins. The aqueous phase was extracted withMTBE (3 L). The organic layer was separated, washed with brine (3×1 L),dried over anhydrous Na₂SO₄, filtered and concentrated below 35° C. togive S-200-N19-3_1 (65 g, crude) as an oil. The crude residue was useddirectly for the next step.

2. To a solution of 2,6-di-tert-butyl-4-methylphenol (340 g, 1.54 mol)in toluene (700 mL) was added drop-wise AlMe₃ (385 mL, 770 mmol, 2 M intoluene) at 0° C. The mixture was stirred at 25° C. for 1 hr and useddirectly as MAD solution. A solution of 200-N19-3_1 (60 g, 220 mmol) inanhydrous toluene (200 mL) and anhydrous DCM (200 mL) was added to MADsolution at −70° C. over a period of 30 mins under N₂. The reactionmixture stirring at −70° C. for 1 h. Then MeMgBr (220 mL, 660 mmol, 3Min ethyl ether) was added drop wise at −70° C. and stirred for 1 h. Thereaction was poured into saturated aqueous citric acid (2 L) at 0° C.and stirring for 30 min, extracted with EtOAc (2×1 L). The combinedorganic phase was washed with saturated brine (2×1 L), dried overanhydrous Na₂SO₄, filtered and concentrated. The residue was purified bysilica gel chromatography (PE/EtOAc=10/1 to 5/1) to afford 200-N19-M22_1(33 g, 52%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.46-5.42 (m, 1H), 2.25-2.40 (m, 1H),2.21-1.60 (m, 13H), 1.35-1.21 (m, 4H), 1.13 (s, 3H), 0.98-0.83 (m, 6H).

3. t-BuOK (31.0 g, 277 mmol) was added in one portion to a suspension ofPh₃PEtBr (102 g, 277 mmol) in Anhydrous THF (500 mL) at 25° C. under N₂.After stirring at 25° C. for 30 min, 200-N19-M22_1 (20 g, 69.3 mmol) wasadded and stirred for 2 h at 25° C. The reaction was quenched withaq.NH₄Cl (800 mL) at 0° C., extracted with EtOAc (2×500 mL). Thecombined organic phase was washed with brine (2×500 mL), dried overNa₂SO₄, filtered and concentrated. The residue was purified by silicagel chromatography (PE/EtOAc=10/1 to 5/1) to afford N-4-14_1 (15 g, 72%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.43-5.40 (m, 1H), 5.16-5.10 (m, 1H),2.41-2.33 (m, 1H), 2.28-1.86 (m, 8H), 1.78-1.71 (m, 1H), 1.69-1.50 (m,11H), 1.41-1.10 (m, 6H), 0.94-0.81 (s, 3H). 4. 9-BNN dimer (66.9 g, 299mmol) was added to a solution of N-4-14_1 (30 g, 99.8 mmol) in anhydrousTHF (500 mL) and stirred for 30 mins at 0° C. under N₂. The reactionmixture was warmed to 50° C. and stirred for 1 hr. After cooling to 0°C. EtOH (100 mL) was added. NaOH.aq (99.8 mL, 5M, 499 mmol) was addedvery slowly. H₂O₂ (53.0 g, 499 mmol, 30% in water) was added slowly andthe inner temperature was maintained below 30° C. The mixture was warmedto 50° C. and stirred for 1 hr. The reaction mixture was cooled andice-water (1 L) was added and stirred 30 min. filtered and concentratedin vacuum to get N-4-11_2 (30 g, crude) as a solid. The crude residuewas used directly for the next step.

5. Silica gel (150 g) and PCC (81.0 g, 376 mmol) were added to asolution of N-4-14_2 (30 g, crude) in DCM (500 mL). The reaction mixturewas warmed to 40° C. and stirred for 1 hr. The reaction mixture wascooled, treated with PE (500 mL), filtered though a pad of silica geland the solid was washed with PE/DCM (500/500 mL). The mother liquor wasfiltered and concentrated under vacuum to get crude product. The residuewas purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1) toafford N-4-14_3 (20 g, impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.43-5.40 (m, 1H), 2.57-2.50 (m, 1H),2.21-2.08 (m, 6H), 1.77-1.43 (m, 10H), 1.37-1.12 (m, 9H), 1.00-0.82 (m,2H), 0.64 (s, 3H).

6. t-BuOK (14.1 g, 126 mmol) was added in one portion to a suspension ofPh₃PMeBr (44.8 g, 126 mmol) in anhydrous THF (300 mL) at 0° C. under N₂.The reaction mixture was stirred at 25° C. for 30 min. N-4-14_3 (20 g,63.1 mmol) was added. The reaction mixture was warmed to 40° C. andstirred for 1 hr. The reaction was poured into ice-water (500 mL) at 0°C. The aqueous phase was extracted with EtOAc (2×400 mL). The combinedorganic phase was washed with brine (2×300 mL), dried over anhydrousNa₂SO₄, filtered and concentrated. The residue was purified by a silicagel column (PE/EtOAc=8/1-5/1) to give N-4-14_4 (19 g, impure) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 5.43-5.40 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H),2.23-2.13 (m, 2H), 1.87-1.64 (m, 11H), 1.42-1.40 (m, 2H), 1.29-1.08 (m,8H), 0.97-0.80 (m, 3H), 0.59 (s, 3H).

7. 9-BNN dimer (40.5 g, 181 mmol) was added in one portion to a solutionof N-4-14_4 (19 g, 60.4 mmol) in anhydrous THF (300 mL) at 0° C. underN₂. The mixture was warmed to 50° C. and stirred for 1 h. The reactionmixture was cooled and EtOH (100 mL) was added. NaOH aq. (60.3 mL, 5M,302 mmol) was added very slowly. H₂O₂ (34.0 g, 302 mmol, 30% in water)was added slowly and the inner temperature was maintained below 10° C.The mixture was warmed to 50° C. and stirred for 1 hr. After cooling,ice-water (1 L) was added and stirred for 30 min. The precipitated solidwas filtered out. The filter cake dried in air to give N-4-11_5 (17 g,crude) as a solid, which was used directly for the next step.

8. To a solution of N-4-14_5 (17 g, crude) in DCM (300 mL) was addedsilica gel (60 g) and PCC (43.9 g, 204 mmol) in one portion at 25° C.The reaction mixture was warmed to 40° C. and stirred for 1 hr. Thereaction mixture was cooled and PE (200 mL) was added. The mixture wasfiltered though a pad of silica gel and the solid was washed with PE/DCM(200/200 mL). filtered and concentrated in vacuum to yield a solid. Theresidue was purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1)to afford N-4-14_6 (5.5 g, impure) as a solid. The residue wasre-crystallized from MeCN (50 mL) at 82° C. to get N-4-14_6 (5 g, 91%)as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.60-9.56 (m, 1H), 5.42-5.40 (m, 1H),2.58-2.51 (m, 1H), 2.40-1.85 (m, 9H), 1.44-1.04 (m, 16H), 1.00-0.80 (m,3H), 0.75-0.71 (m, 3H).

9. To a suspension of Mg (3.96 g, 165 mmol) and 12 (1 mg) in anhydrousTHF (20 mL) was added solution of 1 (12.5 g, 82.7 mmol) in anhydrous THF(63 mL) drop-wise under N₂ at 25° C. and inner temperature was raised to65° C. and stirred for 2 hrs. The mixture was used in the next stepdirectly. Isopentylmagnesium bromide (83.0 mL, 1 M in THF) was added inone portion to a solution of N-4-14_6 (5 g, 15.1 mmol) in anhydrous THF(50 mL) was added at 0° C. under N₂. The reaction mixture was warmed to15° C. and stirred for 1 hr. To the reaction mixture was added saturatedaqueous NH₄Cl (100 mL) solution. The aqueous phase was extracted withEtOAc (3×100 mL). The combined organic phase was washed with saturatedbrine (2×200 mL), dried over anhydrous Na₂SO₄, filtered and concentratedin vacuum to give a crude product. The crude product was purified bysilica gel column (0˜20% of EtOAc in PE) to give N-4-14_7 (2.5 g,impure) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.40-5.37 (m, 1H), 3.67-3.61 (m, 1H),2.19-1.71 (m, 9H), 1.64-1.28 (m, 13H), 1.18-1.03 (m, 7H), 0.95-0.78 (m,12H), 0.69 (s, 3H).

10. To a solution of DMP (10.5 g, 24.8 mmol) was added N-4-14_7 (2.5 g,6.20 mmol) in DCM (40 mL) at 25° C. The reaction mixture was warmed to40° C. and stirred for 1 h. The reaction mixture was quenched withsaturated NaHCO₃ aqueous pH 7˜8 at below 10° C. The Suspension wasfiltered. The DCM phase in filtrate was separated and washed withsaturated NaHCO₃/Na₂S₂O₃ aqueous (1:1, 2×30 mL), the combined organicphase was washed with saturated brine (2×30 mL), dried over Na₂SO₄,filtrate and concentrated in vacuum to get a solid. The residue waspurified by flash column (0˜30% of EtOAc in PE) to give N-4-14_70 (1.5g, 60%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 2.56-2.31 (m, 3H),2.19-1.83 (m, 5H), 1.84-1.42 (m, 12H), 1.30-0.97 (m, 12H), 0.96-0.77 (m,8H), 0.74-0.70 (m, 3H).

11. To a solution of N-4-14_70 (1.5 g, 3.74) in MeOH (10 mL) was slowedadded NaBH₄ (1.42 g, 37.4 mmol) at 25° C. and stirred for 2 hrs.Ice-water (100 mL) was added and the mixture was stirred for 30 mins.The aqueous phase was extracted with DCM (2×20 mL). The combined organicphase was washed with saturated brine (2×20 mL), dried over anhydrousNa₂SO₄, filtered and concentrated in vacuum to obtain a solid. Theresidue was purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1)to afford N-4-14_7 (1 g, 67%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 3.65-3.63 (m, 1H),2.20-2.16 (m, 1H), 2.11-1.88 (m, 5H), 1.86-1.54 (m, 10H), 1.33-0.99 (m,14H), 0.95-0.79 (m, 11H), 0.70 (s, 3H).

SFC Peak 1: Rt=4.644 min and Peak 2 Rt=5.240 min in 10 minchromatography, AD_3_EtOH_DEA_5_40_25ML (““Column: Chiralpak AD-3150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA)Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5%of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.″).

12. The N-4-14_7 (1 g, 2.48 mmol) was purified by SFC (Column: AD (250mm*30 mm, 5 um), Condition: 0.1% NH₃H₂O ETOH, Begin: B: 40%, End B: 40%)to afford 80 (Peak 2, 300 mg, impure) and 83 (Peak 1, 250 mg, impure) asa solid. The 80 (300 mg, impure) was re-crystallized from MeCN (4 mL) at82° C. reflux for 1 hr. The stirred mixture was cooled to 25° C. Thesuspension was filtered under vacuum to get 80 (150 mg, 15%) as a solid.The 83 (250 mg, impure) was re-crystallized from MeCN (3 mL) at 82° C.reflux for 1 hr. The stirred mixture was cooled to 25° C. The suspensionwas filtered under vacuum to provide 83 (150 mg, 15%) as a solid.

83:

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 3.62-3.60 (m, 1H),2.22-1.89 (m, 6H), 1.64-1.49 (m, 9H), 1.46-1.11 (m, 16H), 0.98-0.86 (m,10H), 0.70 (s, 3H).

LCMS Rt=1.268 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. for C₂₇H₄₂ [M+H-2H₂O]⁺ 367, found 367.

SFC Rt=4.609 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

80:

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 3.63-3.62 (m, 1H),2.22-1.67 (m, 10H), 1.64-1.36 (m, 12H), 1.16-1.03 (m, 8H), 0.98-0.80 (m,11H), 0.70 (s, 3H).

LCMS Rt=1.276 min in 2.0 min chromatography, 30-90 AB, purity 99%, MSESI calcd. for C₂₇H₄₂ [M+H-2H₂O]⁺ 367, found 367.

SFC Rt=5.236 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

13. Dry Pd(OH)₂ (200 mg) was added to a solution of 80 (150 mg, 0.372mmol) in THF (3 mL) and MeOH (3 mL) under Ar. The suspension wasdegassed under vacuum and purged with H₂ three times. The mixture wasstirred under H₂ (50 psi) at 50° C. for 12 h to give a black suspension.The reaction mixture was filtered through a pad of Celite and washedwith DCM (3×50 mL). The filtrate was concentrated under vacuum toprovide an oil. The residue was purified by silica gel chromatography(PE/EtOAc=8/1 to 5/1) to afford 74 (20 mg, 13%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.63-3.61 (m, 1H), 1.96-1.85 (m, 2H),1.78-1.50 (m, 8H), 1.45-1.19 (s, 12H), 1.17-1.00 (m, 11H), 0.98-0.83 (m,11H), 0.72-0.59 (m, 3H).

LCMS Rt=1.333 min in 2.0 min chromatography, 30-90 AB, purity 99%, MSESI calcd. for C₂₇H₄₄ [M+H-2H₂O]⁺ 369, found 369.

Example 75: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(75)

1. DMP (2.44 g, 5.76 mmol) was added to a solution of 15_3a (1 g, 2.88mmol) in DCM (10 mL). After that, the reaction was stirred at 25° C. for10 min. The reaction mixture was quenched by adding aqueous saturatedNaHCO₃ (20 mL) solution and aqueous saturated Na₂S₂O₃ (20 mL) solution,extracted with DCM (2×50 mL). The combined organic layer was washed withaqueous saturated NaHCO₃ (3×50 mL) solution and brine (50 mL), driedover Na₂SO₄, filtered and concentrated in vacuum to give S-500-2-9_1 (1g, crude) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.57 (br. s, 1H), 5.35-5.25 (m, 1H), 2.50-2.30(m, 2H), 2.05-1.95 (m, 3H), 1.95-1.80 (m, 1H), 1.75-1.65 (m, 1H),1.65-1.60 (m, 3H), 1.55-1.50 (m, 2H), 1.50-1.40 (m, 2H), 1.40-1.30 (m,1H), 1.25-1.20 (m, 2H), 1.20-1.15 (m, 2H), 1.15-1.10 (m, 6H), 1.05-0.95(m, 5H), 0.90-0.70 (m, 1H), 0.68 (s, 3H).

2. A mixture of magnesium (641 mg, 26.4 mmol) and I₂ (33.5 mg, 0.132mmol) was stirred at 60° C. and a solution of isopentylmagnesium bromide(2 g, 13.2 mmol) in THF (20 mL) was added dropwise under N₂. After that,the reaction mixture was stirred at 60° C. for 1 h. The reaction mixturewas used directly as isopentylmagnesium bromide solution without anypurification. The Grignard solution was added to a solution ofS-500-2-9_1 (1 g, 2.90 mmol) in THF (10 mL) at 0° C. under N₂. Afterthat, the reaction mixture was stirred at 25° C. for 1 h. The reactionmixture was added saturated aqueous NH₄Cl (50 mL) solution, extractedwith EtOAc (2×50 mL), washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated in vacuum to give a crude product. The crudeproduct was purified by silica gel column (EtOAc/PE=¼) to give impureS-500-2-9_2 (560 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.28-5.25 (m, 1H), 3.90-3.80 (m, 0.25H),3.68-3.58 (m, 0.75H), 2.48-2.36 (m, 1H), 2.05-1.95 (m, 3H), 1.95-1.80(m, 1H), 1.80-1.75 (m, 1H), 1.75-1.52 (m, 6H) 1.52-1.42 (m, 6H),1.42-1.32 (m, 3H), 1.32-1.22 (m, 3H), 1.22-1.12 (m, 3H), 1.12-1.02 (m,2H), 1.01 (s, 3H), 1.00-0.92 (m, 1H), 0.92-0.85 (m, 9H), 0.85-0.77 (m,1H), 0.69 (s, 3H).

3. S-500-2-9_2 (560 mg) was purified by SFC (Column: Chiralcel OD-3150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA)Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5%of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.) to giveimpure 30 (160 mg) as a solid and 75 (265 mg, 47%) as a solid.

75:

¹H NMR (400 MHz, CDCl₃) δ 5.35-5.30 (m, 1H), 3.70-3.60 (m, 1H),2.50-2.40 (m, 1H), 2.05-1.90 (m, 4H), 1.85-1.75 (m, 2H), 1.75-1.60 (m,1H), 1.55-1.45 (m, 8H), 1.45-1.25 (m, 8H), 1.25-1.10 (m, 4H), 1.10-1.05(m, 2H), 1.02 (s, 3H), 0.99-0.91 (m, 3H), 0.91-0.89 (m, 4H), 0.88 (s,3H), 0.69 (s, 3H).

LCMS Rt=1.162 min in 1.5 min chromatography, 5-95 AB, purity 99%, MS ESIcalcd. for C₂₈H₄₅[M+H-2H₂O]⁺ 381, found 381.

Example 76: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((S)-1-(1-hydroxycyclopropyl)ethyl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(76)

1. NaClO₂ (374 mg, 4.14 mmol), TEMPO (645 mg, 4.14 mmol) and NaClO (10mL, 10% in water) were added to a solution of N-8-7_1 (500 mg, 1.38mmol) in MeCN (30 mL). After the mixture was stirred at 50° C. for 48hrs, a solid appeared. The solid was collected by filtration andtrituration with DCM (5 mL) to give N-8-26_1 (180 mg, 34%) as a solid.

¹H NMR (400 MHz, MeOD) δ 2.30-2.20 (m, 1H), 1.98-1.91 (m, 1H), 1.86-1.72(m, 1H), 1.71-1.63 (m, 1H), 1.62-1.49 (m, 8H), 1.43-1.31 (m, 4H),1.30-1.20 (m, 4H), 1.19-1.07 (m, 8H), 1.06-0.86 (m, 8H), 0.75-0.65 (m,4H).

2. K₂CO₃ (328 mg, 2.38 mmol) and MeI (686 mg, 4.77 mmol) were added to asolution of N-8-26_1 (180 mg, 0.477 mmol) in DMF (5 mL). After stirringat 20° C. for 16 hrs, the mixture was quenched with 50% NH₄Cl (20 mL)and extracted with EtOAc (3×10 mL). The combined organic phase waswashed with LiCl (3% in water, 30 mL), dried over Na₂SO₄, filtered,concentrated and purified by combi-flash (0-10% of EtOAc in PE) to giveN-8-26_2 (160 mg, 86%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.63 (s, 3H), 2.45-2.36 (m, 1H), 1.92-1.85 (m,1H), 1.74-1.58 (m, 6H), 1.56-1.46 (m, 4H), 1.42-1.19 (m, 9H), 1.18-1.15(m, 3H), 1.13-0.93 (m, 4H), 0.91-0.84 (m, 4H), 0.82 (s, 3H), 0.70-0.61(m, 4H).

3. Ti(i-PrO)₄ (57.9 mg, 0.204 mmol) and EtMgBr (0.204 mL, 3 M in Et₂O,0.612 mmol) were added to a solution of N-8-26_2 (80 mg, 0.204 mmol) inTHF (2 mL) at 20° C. After stirring at 20° C. for 30 minutes, thereaction mixture was quenched with saturated NH₄Cl (30 mL) solution andextracted with EtOAc (3×20 mL). The combined organic layer was washedwith brine (50 mL), dried over Na₂SO₄, filtered and concentrated invacuum to give a crude product, which was purified by silica gel column(0-10% of EtOAc in PE) to afford 76 (16 mg, 20%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 1.98-1.86 (m, 2H), 1.69-1.58 (m, 6H),1.54-1.44 (m, 3H), 1.44-1.29 (m, 4H), 1.28-1.18 (m, 4H), 1.18-1.10 (m,5H), 1.09-0.93 (m, 4H), 0.91-0.81 (m, 9H), 0.71-0.57 (m, 6H), 0.31-0.24(m, 1H).

LCMS Rt=1.184 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₂₆H₄₁ [M+H]⁺ 353, found 353.

Example 77: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(77)

1. A solution of 1-bromo-3-methylbutane (11.7 g, 78 mmol) in THF (8 mL)was added dropwise to a suspension of Mg (4.35 g, 179 mmol) and I₂ (20mg) in THF (2 mL) at 60° C. The mixture was stirred at 60° C. for 1 hr.The mixture was diluted with THF (10 mL) and used directly. Freshlyprepared isopentylmagnesium bromide (19.5 mL, 3.9 M in THF, 76 mmol) wasadded to a solution of S-200-INT_5E (1.0 g, 2.78 mmol) in THF (5 mL)under N₂ at 0° C. The mixture was stirred at 0° C. for 1 hr. NH₄Cl (20mL, sat. aq.) was added to the mixture. The mixture was extracted withEtOAc (2×30 mL). The combined organic phase was washed with brine (100mL), dried over Na₂SO₄, concentrated under vacuum, purified by silicagel (PE/EtOAc=20/1 to 10/1), and re-crystallized from CH₃CN (10 mL) togive 77 (255 mg, 21%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.26 (m, 1H), 3.66-3.59 (m, 1H),2.42-2.32 (m, 1H), 2.07-1.85 (m, 4H), 1.77-1.58 (m, 4H), 1.55-1.38 (m,10H), 1.38-1.19 (m, 5H), 1.19-1.00 (m, 8H), 1.00-0.81 (m, 13H), 0.69 (s,3H).

LCMS Rt=1.306 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. for C₂₉H₄₉O [M+H-H₂O]⁺ 413, found 413.

Example 78: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((1S,2S)-1-cyclopentyl-1-hydroxypropan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(78)

1. NaBH₄ (550 mg, 14.5 mmol) was added to a mixture of N-8-15_2 (240 mg,0.559 mmol) in MeOH (3 mL) and THF (2 mL). The mixture was stirred at15° C. for 0.5 h. Another batch of NaBH₄ (550 mg, 14.5 mmol) was added.The reaction mixture was stirred for another 1 hr. To the reactionmixture was added water (5 mL). The resulting mixture was extracted withEtOAc (2×10 mL). The combined organic layer was washed with brine (10mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column (0˜5% EtOAc in PE) to give and 58 (7 mg, 5%) asa solid and 78 (50 mg, impure) was further purified by flash column(0˜5% of EtOAc in PE) to give 78 (17 mg, 12%) as a solid.

78:

¹H NMR (400 MHz, CDCl₃) δ 3.38-3.47 (m, 1H), 2.01-1.82 (m, 4H),1.71-1.53 (m, 11H), 1.53-1.48 (m, 4H), 1.48-1.30 (m, 5H), 1.30-1.11 (m,7H), 1.11-0.98 (m, 5H), 0.98-0.85 (m, 7H), 0.85-0.80 (m, 3H), 0.65 (s,3H).

LCMS Rt=1.358 min in 2 min chromatography, 30-90AB_7MIN_E, purity 100%,MS ESI calcd. for C₂₉H₄₇ [M+H-2H₂O]⁺ 395, found 395.

HPLC Rt=6.093 min in 10 min chromatography, 50-100AB_10 MIN.M, purity98%.

Example 79: Synthesis of(1R,3S,4S)-4-((3S,5S,8R,9S,10S,13S,14S,17R)-3-hydroxy-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-1-phenylpentane-1,3-diol(79)

The synthesis of 79 is described in Example 13.

79:

¹H NMR (400 MHz, CDCl₃) δ 7.43-7.28 (m, 5H), 5.05-4.94 (m, 1H),4.04-3.91 (m, 1H), 2.51 (brs, 1H), 2.07-1.78 (m, 6H), 1.70-1.61 (m, 4H),1.51-1.41 (m, 3H), 1.39-1.12 (m, 11H), 1.05-0.98 (m, 2H), 0.91-0.81 (m,7H), 0.71-0.60 (m, 4H).

LCMS Rt=1.298 min in 2 min chromatography, 10-80AB_2MIN_E, purity 96.7%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.483 min in 10 min chromatography, IC-3_MeOH(DEA)_40_2.5ML, 100%de.

13:

¹H NMR (400 MHz, CDCl₃) δ 7.40-7.28 (m, 5H), 5.12-5.07 (m, 1H),3.95-3.88 (m, 1H), 2.76 (brs, 1H), 2.08-1.78 (m, 6H), 1.75-1.60 (m, 5H),1.51-1.38 (m, 4H), 1.36-1.09 (m, 9H), 1.00-0.89 (m, 6H), 0.83 (s, 3H),0.71-0.64 (m, 1H), 0.63 (s, 3H).

LCMS Rt=1.309 min in 2 min chromatography, 10-80AB_2MIN_E, purity 100%,MS ESI calcd. for C₃₁H₄₅F₃O₃Na [M+Na]⁺ 545, found 545.

SFC Rt=1.683 min in 5 min chromatography, IC-3_MeOH(DEA)_40_2.5ML,98.94% de.

SFC Rt=4.785 min in 8 min chromatography, AD_MEOH(DEA)_5_40_2,8ML_8MIN,94.03% de.

Example 80: Synthesis of(3S,8R,9S,10R,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(80)

1. The N-4-14_7 (1 g, 2.48 mmol) was purified by SFC (Column: AD (250mm*30 mm, 5 um), Condition: 0.1% NH₃H₂O ETOH, Begin: B: 40%, End B: 40%)to afford 80 (Peak 2, 300 mg, impure) and 83 (Peak 1, 250 mg, impure) asa solid. 80 (300 mg, impure) was re-crystallized from MeCN (4 mL) at 82°C. reflux for 1 hr. The mixture stirred was cooled to 25° C. Thesuspension was filtration in vacuum to get 80 (150 mg, 15%) as a solid.The 83 (250 mg, impure) was re-crystallized from MeCN (3 mL) at 82° C.reflux for 1 hr. The mixture stirred was cool to 25° C. The suspensionwas filtration in vacuum to get 83 (150 mg, 15%) as a solid.

80:

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 3.63-3.62 (m, 1H),2.22-1.67 (m, 10H), 1.64-1.36 (m, 12H), 1.16-1.03 (m, 8H), 0.98-0.80 (m,11H), 0.70 (s, 3H).

LCMS Rt=1.276 min in 2.0 min chromatography, 30-90 AB, purity 99%, MSESI calcd. for C₂₇H₄₂ [M+H-2H₂O]⁺ 367, found 367.

SFC Rt=5.236 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

Example 81: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-ethyl-10,13-dimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(81)

1. S-500-6-1_2 (350 mg) was purified by SFC (Column: AD (250 mm*30 mm, 5um), Condition: 0.1% NH₃.H₂O EtOH, Gradient: from 35% to 35%, Flow Rate(ml/min): 60 mL/min, 25° C.) to afford 81 (Peak 1, 130 mg, 37%) and 65(Peak 2, 180 mg, 52%) as a white solid.

81:

¹H NMR (400 MHz, CDCl3) δ 5.34-5.24 (m, 1H), 4.09-4.00 (m, 1H),2.43-2.33 (m, 1H), 2.14 (d, J=4 Hz, 1H), 2.07-1.80 (m, 5H), 1.77-1.55(m, 5H), 1.53-1.30 (m, 7H), 1.28-1.00 (m, 11H), 1.00-0.91 (m, 1H), 0.85(t, J=8 Hz, 3H), 0.70 (s, 3H).

LCMS Rt=1.220 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. For C₂₅H₃₈F₃O [M+H-H₂O]⁺ 411, found 411.

SFC_E1 Peak 1: Rt=4.561 min in 10 min chromatography,AD_3_EtOH_DEA_5_40_25ML (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 umMobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% ofB in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate:2.5 mL/min Column temp.: 35° C.″), 100% de.

Example 82: Synthesis of(3S,5R,8R,9R,10S,13S,14S,17R)-3-ethyl-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-13-methylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(82)

1. Pd/C (dry, 200 mg) was added to a solution of 44 (200 mg, 0.480 mmol)in MeOH/THF (10 mL/10 mL) under Ar. The suspension was degassed undervacuum and purged with H₂ three times. The mixture was stirred under H₂(50 psi) at 50° C. for 48 hrs to give a black suspension. The reactionmixture was filtered through a pad of Celite and washed with THF (100mL). The filtrate was concentrated to give 28 (30 mg, 15%) as a solidand 82 (30 mg, 15%) as a solid.

82:

¹H NMR (400 MHz, CDCl3) δ 3.63-3.61 (m, 1H), 2.13-2.00 (m, 1H),1.99-1.81 (m, 2H), 1.72-1.57 (m, 6H), 1.54-1.34 (m, 11H), 1.33-1.16 (m,7H), 1.15-0.96 (m, 5H), 0.92-0.85 (m, 13H), 0.81-0.69 (m, 1H), 0.67 (s,3H).

LCMS Rt=1.348 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESIcalcd. For C₂₈H₄₇ [M+H-2H₂O]+ 383, found 383.

Example 83: Synthesis of(3S,8R,9S,10R,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(83)

1. The N-4-14_7 (1 g, 2.48 mmol) was purified by SFC (Column: AD (250mm*30 mm, 5 um), Condition: 0.1% NH₃H₂O ETOH, Begin: B: 40%, End B: 40%)to afford 80 (Peak 2, 300 mg, impure) and 83 (Peak 1, 250 mg, impure) asa solid. The 80 (300 mg, impure) was re-crystallized from MeCN (4 mL) at82° C. reflux for 1 hr. The mixture stirred was cooled to 25° C. Thesuspension was filtered under vacuum to yield 80 (150 mg, 15%) as asolid. The 83 (250 mg, impure) was re-crystallized from MeCN (3 mL) at82° C. reflux for 1 h. The mixture stirred was cool to 25° C. Thesuspension was filtered under vacuum to yield 83 (150 mg, 15%) as asolid.

83:

¹H NMR (400 MHz, CDCl₃) δ 5.41-5.39 (m, 1H), 3.62-3.60 (m, 1H),2.22-1.89 (m, 6H), 1.64-1.49 (m, 9H), 1.46-1.11 (m, 16H), 0.98-0.86 (m,10H), 0.70 (s, 3H).

LCMS Rt=1.268 min in 2.0 min chromatography, 30-90 AB, purity 100%, MSESI calcd. for C₂₇H₄₂ [M+H-2H₂O]⁺ 367, found 367.

SFC Rt=4.609 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100%de.

Example 84: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(84)

1. Pd(OH)₂ (160 mg, dry) was added to a solution of 32 (80 mg, 0.18mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 67 (10 mg, 12%) and 84 (30mg, 37%) as a solid.

84:

¹H NMR (400 MHz, CDCl₃) δ 3.75-3.66 (m, 1H), 2.00-1.90 (m, 2H),1.86-1.75 (m, 2H), 1.73-1.55 (m, 11H), 1.53-1.26 (m, 9H), 1.25-1.15 (m,6H), 1.14-1.03 (m, 5H), 1.02-0.92 (m, 3H), 0.91-0.85 (m, 6H), 0.82 (s,3H), 0.72-0.58 (m, 4H).

LCMS Rt=1.518 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₀H₄₉ [M+H-H₂O]⁺ 409, found 409.

Example 85: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(85)

1. NaBH₄ (0.96 g, 25.4 mmol) was added in portions to a solution ofN-4-1_8 (0.6 g, 1.27 mmol) in THF (10 mL) and MeOH (5 mL) at 15° C. Themixture was stirred at 15° C. for 30 mins. To the mixture was addedNH₄Cl (50 mL, 10%). The mixture was extracted with EtOAc (2×50 mL). Thecombined organic layer was dried over Na₂SO₄, filtered and concentratedunder vacuum and purified by flash column (0˜15% EtOAc in PE) to giveimpure 42 and 85. 42 was triturated from MeCN (10 mL) at 15° C. anddried in vacuum to give 42 (153 mg, 25%) as a solid. 85 was purified byflash column (0˜15% EtOAc in PE) to provide an oil, which was treatedwith MeCN (5 mL) and water (5 mL), concentrated in vacuum to give 85 (70mg, 12%) as a solid.

85:

¹H NMR (400 MHz, CDCl₃) δ 3.66-3.55 (m, 1H), 2.10-1.91 (m, 3H),1.88-1.78 (m, 1H), 1.72-1.55 (m, 6H), 1.50-1.38 (m, 9H), 1.37-0.95 (m,10H), 0.94-0.79 (m, 13H), 0.75-0.61 (m, 4H).

LCMS Rt=1.343 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₈H₄₆F₃O [M+H-H₂O]⁺ 455, found 455.

HPLC Rt=5.14 min in 10.0 min chromatography, 50-100_AB_E, purity 98.56%.

Example 86: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((1S,2S)-1-hydroxy-1-(tetrahydro-2H-pyran-4-yl)propan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(86)

1. A solution of 4-chlorotetrahydro-2H-pyran (1.2 g, 10 mmol) in THF (5mL) was added dropwise to a mixture of Mg (486 mg, 20 mmol) and I₂ (1mg) at 70° C. The mixture was stirred at 50° C. for 0.5 h, diluted withTHF (5 mL) and used directly. To a solution of(tetrahydro-2H-pyran-4-yl)magnesium chloride (4.14 mL, 1 M in THF) wasadded N-8-7_1 (500 mg, 1.38 mmol) in THF (5 mL) at 0° C. under N₂. Thenthe mixture was stirred at 15° C. for another 18 hrs. The reactionmixture was quenched with sat. NH₄Cl (5 mL), and the resulting mixturewas extracted with EtOAc (2×10 mL). The combined organic layer waswashed with brine (5 mL), dried over Na₂SO₄, filtered and concentrated.The residue was purified by flash column (0-15% EtOAc in PE) to giveN-8-17_1 (350 mg, 57%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.08-3.91 (m, 2H), 3.43-3.31 (m, 2H),3.31-3.25 (m, 1H), 1.98-1.91 (m, 2H), 1.91-1.80 (m, 1H), 1.70-1.50 (m,10H), 1.50-1.41 (m, 2H), 1.41-1.32 (m, 5H), 1.32-1.15 (m, 9H), 1.15-0.92(m, 6H), 0.92-0.83 (m, 7H), 0.65 (s, 3H).

2. DMP (0.852 g, 2.01 mmol) was added to a solution of N-8-17_1 (300 mg,0.671 mmol) in DCM (5 mL). After stirring at 15° C. for 10 min, thereaction mixture was quenched with saturated NaHCO₃ (20 mL) until pH ofthe aqueous layer became about 9. The mixture was filtered. The DCMlayer was separated and the aqueous phase was extracted with DCM (2×20mL). The combined organic phase was washed with saturated Na₂S₂O₃aqueous (3×20 mL), sat. NaHCO₃ (20 mL), brine (50 mL), dried overNa₂SO₄, filtered, concentrated and purified by combi-flash (0-30% ofEtOAc in PE) to give N-8-17_3 (200 mg) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 4.08-3.91 (m, 2H), 3.50-3.31 (m, 2H),2.73-2.51 (m, 2H), 1.98-1.79 (m, 1H), 1.79-1.42 (m, 16H), 1.42-1.18 (m,7H), 1.18-0.93 (m, 8H), 0.93-0.79 (m, 6H), 0.68 (s, 4H).

3. LiAlH₄ (50.9 mg, 1.34 mmol) was added to a mixture of N-8-17_3 (200mg, 0.449 mmol) in THF (5 mL) at 0° C. After stirred at 15° C. for 0.5h, the reaction mixture was quenched with water (3 mL) and extractedwith EtOAc (2×10 mL). The combined organic layer was washed with brine(10 mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column (0˜5% EtOAc in PE) to give 86 (23 mg, 11%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 4.08-3.91 (m, 2H), 3.41-3.31 (m, 2H),3.31-3.22 (m, 1H), 2.01-1.79 (m, 3H), 1.70-1.61 (m, 1H), 1.61-1.53 (m,8H), 1.53-1.51 (m, 1H), 1.51-1.39 (m, 5H), 1.39-1.13 (m, 8H), 1.13-0.92(m, 5H), 0.92-0.85 (m, 7H), 0.82 (s, 3H), 0.66 (s, 4H).

LCMS Rt=4.832 min in 7 min chromatography, 30-90AB_7MIN_E, purity 100%,MS ESI calcd. for C₂₉H₄₇O [M+H-2H₂O]⁺ 411, found 411.

HPLC Rt=6.338 min in 10 min chromatography, 30-90AB_1.2 mL e. Met, 100%purity.

Example 87: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3R)-4-(4,4-dimethylcyclohexyl)-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(87)

1. Pd(OH)₂ (150 mg, dry) was added to a solution of 12 (100 mg, 0.206mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50Psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 87 (12 mg, 12%) and 20 (11mg, 11%) as a solid.

87:

¹H NMR (400 MHz, CDCl₃) δ 3.82-3.75 (m, 1H), 2.00-1.83 (m, 2H),1.80-1.58 (m, 7H), 1.52-1.42 (m, 4H), 1.40-1.27 (m, 10H), 1.25-1.14 (m,10H), 1.13-0.98 (m, 6H), 0.96 (s, 3H), 0.94-0.82 (m, 12H), 0.67 (s, 3H).

LCMS Rt=1.734 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₃H₅₅ [M+H-2H₂O]⁺ 451, found 451.

Example 88: Synthesis of(3S,5S,8R,9R,10S,13S,14S,17R)-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(88)

1. Pd(OH)₂ (200 mg) was to a solution of 83 (150 mg, 0.372 mmol) in MeOH(3 mL) and THF (3 mL) was added under Ar. The suspension was degassedunder vacuum and purged with H₂ three times. The mixture was stirredunder H₂ (50 psi) at 50° C. for 12 h to give a black suspension. Thereaction mixture was filtered through a pad of Celite and washed withDCM (3×50 mL). The filtrate was concentrated under vacuum to provide anoil. The residue was purified by silica gel chromatography (PE/EtOAc=8/1to 5/1) to afford 88 (18 mg, 12%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 3.62-3.59 (m, 1H), 1.97-1.81 (m, 2H),1.76-1.50 (m, 12H), 1.46-1.28 (m, 5H), 1.24-1.03 (m, 12H), 0.95-0.83 (m,11H), 0.74-0.57 (m, 5H).

LCMS Rt=1.317 min in 2.0 min chromatography, 30-90 AB, purity 99%, MSESI calcd. for C₂₇H₄₄ [M+H-2H₂O]⁺ 369, found 369.

Example 89: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((1S,2S)-1-hydroxy-1-(pyridin-3-yl)propan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(89)

1. N-8-19_1 (3340) (100 mg, 0.227 mmol) was separated by SFC (column: AD(250 mm*30 mm, 5 um), gradient: 50-50% B (A=0.05% NH₃/H₂O, B=MeOH), flowrate: 80 mL/min) to give 7 (Peak 1, 57 mg, 57%) and 89 (Peak 2, 8 mg,8%) as a solid.

SFC Peak 1: Rt=1.798 min and Peak 2 Rt=1.985 min in 3 minchromatography, AD-H_3UM_4_5_40_4ML (“Chiralpak AD-3 50*4.6 mm I.D., 3um Mobile phase: A: CO2 B: iso-propanol (0.05% DEA) Gradient: from 5% to40% of B in 1.4 min and hold 40% for 1.05 min, then 5% of B for 0.35 minFlow rate: 4 mL/min Column temp.: 40° C.″).

Example 90: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((1R,2S)-1-cyclopropyl-1-hydroxypropan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(90)

1. NaBH₄ (1.18 g, 17.4 mmol) was added five times, every five minutes,to a solution of N-8-13_2 (140 mg, 0.347 mmol) in MeOH (1 mL) and THF (1mL). The mixture was stirred at 15° C. for 30 minutes. The mixture wasquenched with sat.NH₄Cl (50 mL) and extracted with EtOAc (3×20 mL). Thecombined organic phase was dried over Na₂SO₄, filtered, concentrated andpurified by combi-flash (25% of EtOAc in PE) to give 10 (26 mg, 19%) asa solid and 90 (12 mg, 9%) as a solid.

90:

¹H NMR (400 MHz, CDCl₃) δ 3.00-2.80 (m, 1H), 2.01-1.95 (m, 1H),1.75-1.60 (m, 5H), 1.47-1.18 (m, 13H), 1.15-0.79 (m, 18H), 0.70-0.60 (m,4H), 0.58-0.50 (m, 1H), 0.48-0.40 (m, 1H), 0.38-0.30 (m, 1H), 0.24-0.16(m, 1H).

LCMS Rt=3.796 min in 7.0 min chromatography, 30-90AB_7MIN_E, purity100%, MS ESI calcd. for C₂₇H₄₃ [M+H-2H₂O]⁺ 367, found 367.

HPLC Rt=13.689 min in 30 min chromatography, 70-90AB_1_30MIN.M, purity98%.

Example 91: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxybutan-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(91)

The synthesis of 91 is described in Example 13.

91:

¹H NMR (400 MHz, CDCl₃) δ 3.98-3.88 (m, 1H), 2.11-2.02 (m, 1H), 2.00 (s,1H), 1.98-1.88 (m, 2H), 1.85-1.79 (m, 1H), 1.73-1.58 (m, 4H), 1.52-1.20(m, 11H), 1.19-1.11 (m, 4H), 1.10-1.00 (m, 3H), 0.97-0.89 (m, 4H), 0.85(s, 3H), 0.75-0.68 (m, 1H), 0.66 (s, 3H).

LCMS Rt=1.155 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, MSESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

HPLC Rt=5.23 min in 10.0 min chromatography, 30-90_AB_E, purity 98.88%,d.e. 100%.

Example 92: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-3-hydroxy-6-methylheptan-2-yl)-3,10,13-trimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(92)

1. DMP (2.42 g, 5.72 mmol) was added to a solution of N-4-16_1 (1.00 g,2.86 mmol) in DCM (20 mL). After that, the reaction was stirred at 15°C. for 10 min. Aqueous saturated NaHCO₃ (20 mL) solution and aqueoussaturated Na₂S₂O₃ (20 mL) solution were added to the reaction mixture,then extracted with DCM (2×20 mL). The combined organic layer was washedwith aqueous saturated NaHCO₃ (3×60 mL) solution and brine (60 mL),dried over Na₂SO₄, filtered and concentrated under vacuum to give asolid. The residue was purified by silica gel chromatography (PE/EtOAc=0to 30%) to afford N-4-16_2 (800 mg, 81%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 9.58-9.53 (m, 1H), 2.41-2.31 (m, 1H),1.96-1.81 (m, 4H), 1.89-1.34 (m, 10H), 1.32-1.21 (m, 10H), 1.16-1.09 (m,5H), 0.97 (s, 3H), 0.89-0.84 (m, 1H), 0.69 (s, 3H).

2. N-4-16_2 (300 mg, 0.86 mmol) in THF (10 mL) was added to a solutionof isopentylmagnesium bromide (4.32 mL, 2 M in ether, 8.65 mmol) at 25°C. under N₂. The mixture was stirred at 25° C. for 30 minutes, quenchedby saturated NH₄Cl (10 mL) and extracted with ethyl acetate (3×10 mL).The organic layer was washed with brine (20 mL), dried over Na₂SO₄ andfiltered, concentrated under vacuum and purified by flash column (0˜15%of EtOAc in PE) to give N-4-16 (200 mg, impure) as a solid, which wastriturated in MeCN (10 mL) at 25° C. to give 92 (141 mg, 70%) as asolid.

¹H NMR (400 MHz, CDCl₃) δ 3.67-3.57 (m, 1H), 2.01-1.77 (m, 4H),1.67-1.57 (m, 4H), 1.55-1.26 (m, 14H), 1.25-1.21 (m, 5H), 1.19-0.99 (m,7H), 0.96 (s, 3H), 0.93-0.84 (m, 9H), 0.66 (s, 3H).

LCMS Rt=1.367 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₈H₄₇ [M+H-2H₂O]⁺ 383, found 383.

Example 93: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-4-phenylbutan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(93)

1. A mixture of N-6-5 and N-6-6 (190 mg, 0.420 mmol) was separated byprep. HPLC (Column: YMC-Actus Triart C18 100*30 mm*5 um; condition:water (0.05% HCl)-ACN; Gradient: 90-100% B; Flow rate: 25 mL/min) togive 93 (56 mg, 30%) as a solid and 32 (12 mg, 6%) as a solid.

93:

¹H NMR (400 MHz, CDCl₃) δ 7.34-7.28 (m, 2H), 7.25-7.18 (m, 3H),3.95-3.86 (m, 1H), 2.87-2.75 (m, 1H), 2.69-2.58 (m, 1H), 2.01-1.91 (m,1H), 1.90-1.79 (m, 1H), 1.69-1.58 (m, 4H), 1.55-1.41 (m, 6H), 1.40-1.11(m, 11H), 1.07-0.95 (m, 6H), 0.91-0.80 (m, 7H), 0.69-0.59 (m, 4H).

LCMS Rt=1.334 min in 2.0 min chromatography, 30-90AB, purity 100%, MSESI calcd. for C₃₁H48O₂Na [M+Na]⁺ 475, found 475.

Example 94: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(94)

1. Pd(OH)₂ (300 mg, dry) was added to a solution of 100 (150 mg, 0.338mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 94 (6 mg, 4%) and 98 (46 mg,30%) as a solid.

94:

¹H NMR (400 MHz, CDCl₃) δ 3.78-3.66 (m, 1H), 1.98-1.72 (m, 7H),1.69-1.59 (m, 4H), 1.48-1.32 (m, 12H), 1.27-1.07 (m, 12H), 1.06-1.00 (m,3H), 0.97 (s, 3H), 0.94-0.85 (m, 7H), 0.66 (s, 3H).

LCMS Rt=1.639 min in 2.0 min chromatography, 30-90AB_E, purity 98.8%, MSESI calcd. for C₃₀H₄₉ [M+H-2H₂O]⁺ 409, found 409.

Example 95: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-3-(methoxymethyl)-10,13-dimethyl-17-((2S,3R)-4,4,4-trifluoro-3-hydroxybutan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(95)

1. The N-004-029A (0.45 g, 1.01 mmol) was purified by SFC (Column: AD(250 mm*30 mm, 5 um), Condition: 0.1% NH₃H₂O ETOH, Begin B: 30%, End B:30%) to afford 39 (PK1: 120 mg, 26.7%) as a white solid and 95 (PK2: 200mg, 44.5%) as a white solid.

95:

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.29 (m, 1H), 4.06-3.99 (m, 1H), 3.37 (s,3H), 3.30-3.19 (m, 2H), 2.54 (s, 1H), 2.42-2.33 (m, 1H), 2.17-2.07 (m,1H), 2.20-1.85 (m, 5H), 1.77-1.63 (m, 4H), 1.51-0.83 (m, 17H), 0.73 (s,3H).

LCMS Rt=1.103 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%,MS ESI calcd. for C₂₄H₃₄F₃O [M-CH₅O₂]⁺ 395, found 395.

Example 96: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3-ethyl-17-((2S,3R)-3-hydroxy-6-methylheptan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(96)

1. Pd(OH)₂ (200 mg) was added to a solution of 52 (50 mg, 0.116 mmol) inMeOH (10 mL). The mixture was stirred at 50° C. under H₂ (50 Psi). Themixture was filtered, concentrated and purified by combi-flash (0-10% ofEtOAc in PE) to give 24 (15 mg, 30%) as a solid and 96 (1.2 mg, 3%) as asolid.

96:

¹H NMR (400 MHz, CDCl₃) δ 3.67-3.55 (m, 1H), 2.01-1.83 (m, 2H),1.80-1.62 (m, 4H), 1.61-1.56 (m, 2H), 1.55-1.50 (m, 1H), 1.49-1.31 (m,10H), 1.30-1.10 (m, 11H), 1.09-1.00 (m, 3H), 0.96 (s, 3H), 0.94-0.86 (m,12H), 0.67 (s, 3H).

LCMS t_(R)=1.326 min in 2 min chromatography, 30-90AB_ELSD, purity100.0%, MS ESI calcd. for C₂₉H₄₉ [M+H-2H₂O]⁺ 397, found 397.

Example 97: Synthesis of(3S,5R,8R,9S,10S,13S,14S,17R)-3,10,13-trimethyl-17-((2S,3S)-4,4,4-trifluoro-3-hydroxybutan-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(97)

1. To a solution of N-004-017_3 (100 mg, 0.19 mmol) in THF (2 mL) andMeOH (1 mL) and water (1 mL) was added KOH (53.8 mg, 0.96 mmol). Themixture was stirred at 60° C. for 16 hrs. The mixture was poured intowater (20 mL) and extracted with EtOAc (2×40 mL). The combined organiclayer was washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by flash column (0˜15% of EtOAcin PE) to give 97 (80 mg, impure) as a white solid. 97 (80 mg, 0.19mmol) was purified by flash column (0˜10% of EtOAc in PE) to give 97 (60mg, impure). To a solution of 97 (40 mg, 0.096 mmol) in methanol (3 mL)was added NaBH₄ (10.8 mg, 0.28 mmol) in one portion at 0° C. under N₂.The mixture was stirred at 0° C. for 30 min. The mixture was poured intowater (10 mL) and stirred for 20 min. The aqueous phase was extractedwith EtOAc (3×10 mL). The combined organic phase was washed with brine(2×20 mL), dried over Na₂SO₄, filtered and concentrated to give a crudeproduct, which was combined with another batch from 20 mg of impure 97,the residue was purified by flash column (0˜10% of EtOAc in PE) to give97 (31 mg, 31%) as a white solid.

¹H NMR (400 MHz, CDCl3) δ 4.11-3.96 (m, 1H), 2.18-2.11 (d, J_(ab)=6.4Hz, 1H), 2.02-1.77 (m, 5H), 1.68-1.57 (m, 3H), 1.49-1.24 (m, 11H),1.23-1.19 (m, 5H), 1.18-1.01 (m, 7H), 0.96 (s, 3H), 0.67 (s, 3H).

LCMS Rt=1.124 min in 2 min chromatography, 30-90AB_2MIN_E.M, purity100%, MS ESI calcd. for C₂₄H₃₈F₃O [M+H-H₂O]⁺ 399, found 399.

Example 98: Synthesis of(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(98)

1. Pd(OH)₂ (300 mg, dry) was added to a solution of 100 (150 mg, 0.338mmol) in MeOH (20 mL). The mixture was stirred at 50° C. under H₂ (50psi) for 48 hrs. The mixture was filtered, concentrated and purified bycombi-flash (0-15% of EtOAc in PE) to give 94 (6 mg, 4%) and 98 (46 mg,30%) as a solid.

98:

¹H NMR (400 MHz, CDCl₃) δ 3.78-3.67 (m, 1H), 1.97-1.74 (m, 6H),1.68-1.56 (m, 8H), 1.53-1.45 (m, 4H), 1.44-1.31 (m, 10H), 1.28-1.21 (m,1H), 1.16-0.96 (m, 9H), 0.91-0.85 (m, 6H), 0.82 (s, 3H), 0.69-0.61 (m,4H).

LCMS Rt=1.582 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₀H₄₉ [M+H-2H₂O]⁺ 409, found 409.

Example 99: Synthesis for(3S,5S,8R,9S,10S,13S,14S,17R)-17-((2S,3S,E)-3-hydroxy-5-phenylpent-4-en-2-yl)-10,13-dimethyl-3-(trifluoromethyl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-ol(99)

1. NaBH₄ (419 mg, 11.1 mmol) was added in portions to a solution ofN-003-005_1 (140 mg, 0.278 mmol) in THF (2 mL) and MeOH (1 mL) at 20° C.The mixture was stirred at 20° C. for 10 min. The reaction was quenchedwith water (20 mL) and NH₄Cl (20 mL, sat.). The mixture was extractedwith EtOAc (50 mL). The organic layer was concentrated under vacuum andpurified by prep-TLC (PE/EtOAc=4/1) to give N-003-005 (50 mg, impure)and 14 (50 mg) both as a solid.

The impure 99 (50 mg) was purified by SFC (Instrument: SFC 1; Column: OD(250 mm*30 mm, 5 um); Condition: 0.10/NH₃H₂O EtOH; Begin B: 40%; End B:40%; Flow Rate (mL/min): 50; Injections: 60) to provide a solid whichwas dissolved in MeCN (20 mL) and concentrated in vacuum to give 99 (17mg) as a solid.

99:

¹H NMR (400 MHz, CDCl₃) δ 7.44-7.36 (m, 2H), 7.36-7.28 (m, 2H),7.25-7.20 (m, 1H), 6.58 (d, J=16.0 Hz, 1H), 6.24 (dd, J=4.8, 16.0 Hz,1H), 4.49-4.40 (m, 1H), 2.09-1.91 (m, 4H), 1.86-1.75 (m, 1H), 1.72-1.58(m, 5H), 1.52-1.04 (m, 14H), 0.94 (d, J=6.4 Hz, 3H), 0.91-0.87 (m, 1H),0.86 (s, 3H), 0.75-0.70 (m, 1H), 0.69 (s, 3H).

LCMS Rt=1.280 min in 2 min chromatography, 30-90AB_2MIN_E, purity 98.5%,MS ESI calcd. for C₃₁H₄₂F₃O [M+H-H₂O]⁺ 487, found 487.

HPLC Rt=6.29 min in 8 min chromatography, 30-90_AB_1.2 ml, 100% d.e.

Example 100: Synthesis of(3S,8S,9S,10R,13S,14S,17R)-17-((2S,3S)-4-cyclopentyl-3-hydroxybutan-2-yl)-3-ethyl-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ol(100)

1. A solution of (bromomethyl)cyclopentane (1.8 g, 11.0 mmol) in THF (11mL) was added dropwise to a suspension of Mg (528 mg, 22.0 mmol) and I₂(55.8 mg, 0.22 mmol) in THF (3 mL) at 75° C. The mixture was stirred at75° C. for 1 hr. A mixture of (cyclopentylmethyl)magnesium bromide (11.1mL, 11.1 mmol, 1M in THF) was slowly added to a solution of S-500-6-1_1(800 mg, 2.23 mmol) in THF (30 mL) at 15° C. After addition, the mixturewas stirred at 15° C. for 1 hr. The mixture was quenched with sat. NH₄Cl(40 mL) and extracted with EtOAc (3×20 mL). The combined organic phasewas washed with brine (2×30 mL), dried over Na₂SO₄, filtered andconcentrated and purified by combi-flash (0-15% of EtOAc in PE) to give100 (350 mg, 35%) as a solid.

¹H NMR (400 MHz, CDCl₃) δ 5.32-5.26 (m, 1H), 3.77-3.69 (m, 1H),2.41-2.31 (m, 1H), 2.09-1.89 (m, 4H), 1.88-1.69 (m, 4H), 1.68-1.55 (m,6H), 1.54-1.27 (m, 12H), 1.26-1.15 (m, 2H), 1.14-1.05 (m, 4H), 1.04-0.99(m, 5H), 0.98-0.88 (m, 4H), 0.87-0.81 (m, 3H), 0.69 (s, 3H).

LCMS Rt=5.661 min in 7.0 min chromatography, 30-90AB_E, purity 100%, MSESI calcd. for C₃₀H₄₇ [M+H-2H₂O]⁺ 407, found 407.

TABLE 1 Data for Exemplary Compounds. GluN2A PCA GluN2A GluN2A % PCAEC₅₀ PCA E_(Max) GluN2B PCA G1uN2B PCA potentiation Compound (nM) (%)EC₅₀ (nM) E_(Max) (%) at 1 μM 1 H 2 I 3 C D C D 4 A D C D I 6 C D C D G7 G 8 C D C D I 9 H 10 I 11 I 12 C D C D G 13 A D A E 15 H 16 H 18 G 19H 20 C D C D H 21 H 22 G 23 C D C E H 24 C D C D 25 C D C D 28 C D C E29 C D C D 30 C E C E 31 A E B E 32 C D C D G 36 C D C D 38 C D C D H 39H 40 C D C E G 41 H 43 I 44 C D C D 45 H 46 C D C D H 47 C D C D 48 H 49C D C D 50 H 52 C D C D 53 H 55 H 56 I 57 I 58 H 59 C D C D I 62 H 64 H65 H 67 C D C D G 68 H 69 C D C D G 70 H 71 H 72 C D C D H 73 H 74 H 75C D C D 77 C D C D 79 C D C D 80 H 81 H 82 C D C D 83 H 84 C D C D G 85G 86 H 87 C D C D G 88 H 90 G 91 H 94 C D C D H 95 I 96 B D C D G 97 H98 C D C D H 100 C D C D H For Table 1, “A” indicates an EC₅₀ of 1 to100 nM, “B” indicates an EC₅₀ of greater than 100 nM up to 1 μM, “C”indicates an EC₅₀ greater than 1 μM; “D” indicates an E_(max) of up to100%, “E” indicates an E_(max) between 100% and 500%, “F” indicates anE_(max) greater than 500%; “G” indicates a % potentiation between andincluding 10% and −10%, “H” indicates a % potentiation less than −10%and greater than or equal to −40%, and “I” indicates a % potentiationless than −40%.

Other Embodiments

In the claims articles such as “a,” “an,” and “the” may mean one or morethan one unless indicated to the contrary or otherwise evident from thecontext. Claims or descriptions that include “or” between one or moremembers of a group are considered satisfied if one, more than one, orall of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The invention includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Theinvention includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process.

Furthermore, the invention encompasses all variations, combinations, andpermutations in which one or more limitations, elements, clauses, anddescriptive terms from one or more of the listed claims is introducedinto another claim. For example, any claim that is dependent on anotherclaim can be modified to include one or more limitations found in anyother claim that is dependent on the same base claim. Where elements arepresented as lists, e.g., in Markush group format, each subgroup of theelements is also disclosed, and any element(s) can be removed from thegroup. It should it be understood that, in general, where the invention,or aspects of the invention, is/are referred to as comprising particularelements and/or features, certain embodiments of the invention oraspects of the invention consist, or consist essentially of, suchelements and/or features. For purposes of simplicity, those embodimentshave not been specifically set forth in haec verba herein. It is alsonoted that the terms “comprising” and “containing” are intended to beopen and permits the inclusion of additional elements or steps. Whereranges are given, endpoints are included. Furthermore, unless otherwiseindicated or otherwise evident from the context and understanding of oneof ordinary skill in the art, values that are expressed as ranges canassume any specific value or sub-range within the stated ranges indifferent embodiments of the invention, to the tenth of the unit of thelower limit of the range, unless the context clearly dictates otherwise.

This application refers to various issued patents, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference. If there is a conflict between any ofthe incorporated references and the instant specification, thespecification shall control. In addition, any particular embodiment ofthe present invention that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Because such embodimentsare deemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the invention can be excluded from any claim,for any reason, whether or not related to the existence of prior art.

Those skilled in the art will recognize or be able to ascertain using nomore than routine experimentation many equivalents to the specificembodiments described herein. The scope of the present embodimentsdescribed herein is not intended to be limited to the above Description,but rather is as set forth in the appended claims. Those of ordinaryskill in the art will appreciate that various changes and modificationsto this description may be made without departing from the spirit orscope of the present invention, as defined in the following claims.

What is claimed is:
 1. A compound of Formula (I-63):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is alkyl; R²is alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, orheteroaryl and R³ is hydrogen, alkyl, carbocyclyl, heterocyclyl, aryl,or heteroaryl, or R² and R³, together with the carbon atom to which theyare attached form a 3-8 membered ring; each of R⁴ and R⁵ isindependently hydrogen, halo, or —OR^(C), wherein R^(C) is hydrogen orC₁-C₆alkyl, or R⁴ and R⁵, together with the carbon atom to which theyare attached form an oxo group; R⁶ is absent or hydrogen; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and wherein when both of

are single bonds, R⁶ is hydrogen.
 2. The compound of claim 1, or thepharmaceutically acceptable salt thereof, wherein R¹ is C₁-C₆alkyl. 3.The compound of claim 1, or the pharmaceutically acceptable saltthereof, wherein R² is alkyl.
 4. The compound of claim 1, or thepharmaceutically acceptable salt thereof, wherein R² is unsubstitutedC₁-C₆alkyl or C₁-C₆haloalkyl.
 5. The compound of claim 1, or thepharmaceutically acceptable salt thereof, wherein R² is unsubstitutedC₁-C₆alkyl, C₁-C₆haloalkyl, carbocyclyl, carbocyclylalkyl, aralkyl, orheterocyclylalkyl.
 6. The compound of claim 1, or the pharmaceuticallyacceptable salt thereof, wherein R² is aryl or heteroaryl and R³ ishydrogen.
 7. The compound of claim 1, or the pharmaceutically acceptablesalt thereof, wherein R² is carbocyclyl or heterocyclyl and R³ ishydrogen.
 8. The compound of claim 1, or the pharmaceutically acceptablesalt thereof, wherein R² and R³, together with the carbon atom to whichthey are attached form a 3-8 membered carbocyclic or heterocyclic ring.9. The compound of claim 1, or the pharmaceutically acceptable saltthereof, wherein the compound of Formula (I-63) is selected from thegroup consisting of a compound of Formula (I-A63), Formula (I-B63), andFormula (I-C63):


10. A compound of Formula (I-67):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is alkyl;each of R² and R³ is independently hydrogen, alkyl, carbocyclyl,heterocyclyl, aryl, or heteroaryl, or R² and R³, together with thecarbon atom to which they are attached form a 3-8 membered ring; each ofR⁴ and R⁵ is independently hydrogen, halo, or —OR^(C), wherein R^(C) ishydrogen or C₁-C₆alkyl, or R⁴ and R⁵, together with the carbon atom towhich they are attached form an oxo group; R⁶ is absent or hydrogen; and

represents a single or double bond, wherein when one of

is a double bond, the other

is a single bond and R⁶ is absent; and wherein when both of

are single bonds, R⁶ is hydrogen.
 11. The compound of claim 10, or thepharmaceutically acceptable salt thereof, wherein each of R² and R³ isindependently hydrogen or alkyl.
 12. The compound of claim 10, or thepharmaceutically acceptable salt thereof, wherein each of R² and R³ isindependently hydrogen or C₁-C₆haloalkyl.
 13. The compound of claim 10,or the pharmaceutically acceptable salt thereof, wherein the compound ofFormula (I-67) is selected from the group consisting of a compound ofFormula (I-A67), Formula (I-B67), and Formula (I-C67):


14. A compound, or a pharmaceutically acceptable salt thereof, selectedfrom the group consisting of:


15. A pharmaceutical composition comprising a compound of claim 1, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 16. A method for treating a disorder, the methodcomprising administering to a subject in need thereof an effectiveamount of a compound of claim 1, or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition comprising a compound of claim1, or a pharmaceutically acceptable salt thereof, wherein the disorderis a gastrointestinal (GI) disorder, a structural disorder affecting theGI system, an anal disorder, a colon polyp, cancer, diabetes, a sterolsynthesis disorder, or colitis.
 17. The method according to claim 16,wherein the disorder is a gastrointestinal (GI) disorder, wherein thegastrointestinal (GI) disorder is inflammatory bowel disease.
 18. Amethod for treating a central nervous system (CNS)-related condition,the method comprising administering to a subject in need thereof aneffective amount of a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition comprising acompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein the CNS-related condition is an adjustment disorder, an anxietydisorder, a cognitive disorder, a dissociative disorder, an eatingdisorder, a mood disorder, a psychotic disorder, a sleep disorder, asubstance-related disorder, a personality disorder, an autism spectrumdisorder, a neurodevelopmental disorder, multiple sclerosis, a sterolsynthesis disorder, pain, encephalopathy secondary to a medicalcondition, attention deficit disorder, attention deficit hyperactivitydisorder, a seizure disorder, stroke, traumatic brain injury, a movementdisorder, vision impairment, hearing loss, or tinnitus.
 19. The methodaccording to claim 18, wherein the CNS-related condition is an anxietydisorder, wherein the anxiety disorder is obsessive-compulsive disorder,posttraumatic stress disorder, or social phobia.
 20. The methodaccording to claim 18, wherein the CNS-related condition is a psychoticdisorder, wherein the psychotic disorder is schizophrenia.
 21. Themethod according to claim 18, wherein the CNS-related condition is anautism spectrum disorder.
 22. The method according to claim 18, whereinthe CNS-related condition is attention deficit disorder or attentiondeficit hyperactivity disorder.
 23. The method according to claim 18,wherein the CNS-related condition is a movement disorder, wherein themovement disorder is Huntington's disease or Parkinson's disease. 24.The method according to claim 18, wherein the CNS-related condition is acognitive disorder, wherein the cognitive disorder is Alzheimer'sdisease.
 25. A method for modulating an NMDA receptor, the methodcomprising contacting the NMDA receptor with an effective amount of acompound of claim 1, or a pharmaceutically acceptable salt thereof, or apharmaceutical composition comprising a compound of claim 1, or apharmaceutically acceptable salt thereof.